Preliminary research on tailored fluid therapy in pigs: comparing customized ionic solutions with Hartmann's solution.

BACKGROUND: Fluid therapy in veterinary medicine is pivotal for treating various conditions in pigs; however, standard solutions, such as Hartmann's solution, may not optimally align with pig physiology. This study explored the development and efficacy of a customized fluid therapy tailored to the ionic concentrations of pig blood, aiming to enhance treatment outcomes and safety in both healthy and diseased pigs. RESULTS: The study involved two experiments: the first to assess the safety and stability of customized fluids in healthy pigs, and the second to evaluate the efficacy in pigs with clinical symptoms of dehydration. In healthy pigs, the administration of customized fluids showed no adverse effects, with slight alterations observed in pO2, hematocrit, and glucose levels in some groups. In symptomatic pigs, the customized fluid group did not show any improvement in clinical symptoms, with no significant changes in blood chemistry or metabolite levels compared to controls. The customized fluid group showed a mild increase in some values after administration, yet within normal physiological ranges. The study reported no significant improvements in clinical or dehydration status, attributing the observed variations in blood test results to the limited sample size and anaesthesia effects rather than fluid characteristics. CONCLUSIONS: Customized fluid therapy, tailored to mimic the ionic concentrations of pig blood, appears to be a safe and potentially more effective alternative to conventional solutions such as Hartmann's solution for treating pigs under various health conditions. Further research with larger sample sizes and controlled conditions is recommended to validate these findings and to explore the full potential of customized fluid therapy in veterinary practice.

Surveillance of wild animals carrying infectious agents based on high-throughput screening platform in the Republic of Korea.

BACKGROUND: Infectious diseases transmitted by wild animals are major threats to public health. This study aimed to investigate the potential of rescued wild animals that died of unknown causes as reservoirs of infectious agents. From 2018 to 2019, 121 dead wild animals (55 birds and 66 mammals) were included in this study. All wild animals died during treatment after anthropogenic events. After deaths of animals, necropsies were performed and trachea, lungs, large intestine (including stool), and spleen were collected to determine causes of deaths. A high-throughput screening (HTS) quantitative polymerase chain reaction (qPCR) designed to detect 19 pathogens simultaneously against 48 samples in duplicate was performed using nucleic acids extracted from pooled tissues and peripheral blood samples. If positive, singleplex real-time PCR was performed for individual organs or blood samples. RESULTS: The HTS qPCR showed positive results for Campylobacter jejuni (10/121, 8.3%), Campylobacter coli (1/121, 0.8%), Mycoplasma spp. (78/121, 64.5%), and Plasmodium spp. (7/121, 5.7%). Singleplex real-time PCR confirmed that C. jejuni was detected in the large intestine but not in the blood. C. coli was only detected in the large intestine. Mycoplasma spp. were detected in all organs, having the highest proportion in the large intestine and lowest in the blood. Plasmodium spp. was also detected in all organs, with proportions being were similar among organs. CONCLUSIONS: This study shows that wild animals can become carriers of infectious agents without showing any clinical symptoms.

Application of a modified nictitating membrane flap in a fledgling feral pigeon (Columba livia domestica) with bullous keratopathy.

A fledgling feral pigeon with systemic protozoal infection was referred with corneal protrusion in the right eye after being treated for a corneal ulcer for 12 days. Ophthalmic examination revealed a corneal bulla covering almost the entire central cornea and preventing the eyelids from closing. Gelatinous corneal stroma with numerous heterophils surrounding the corneal bulla was also detected on cytologic examination. Bullous keratopathy and melting keratitis in the right eye were diagnosed. Temporary tarsorrhaphy with topical eye drops was prescribed for a week; however, the bulla persisted. A modified nictitating membrane flap was created under general anesthesia and maintained for 2 weeks. The corneal bulla resolved, and corneal thickness was restored. The pigeon was presented again with recurrence of the corneal bulla in the right eye after 45 days. Alternative surgical options were recommended; however, the pigeon was euthanized because the protozoa-induced intra-oral and intra-aural masses caused malformation of the beaks, which made voluntary feeding impossible.

Analysis of Five Arboviruses and Culicoides Distribution on Cattle Farms in Jeollabuk-do, Korea.

Arthropod-borne viruses (Arboviruses) are transmitted by arthropods such as Culicoides biting midges and cause abortion, stillbirth, and congenital malformation in ruminants, apparently leading to economic losses to farmers. To monitor the distribution of Culicoides and to determine their relationship with different environmental conditions (temperature, humidity, wind speed, and altitude of the farms) on 5 cattle farms, Culicoides were collected during summer season (May-September) in 2016 and 2017, and analyzed for identification of species and detection of arboviruses. About 35% of the Culicoides were collected in July and the collection rate increased with increase in temperature and humidity. The higher altitude where the farms were located, the more Culicoides were collected on inside than outside. In antigen test of Culicoides against 5 arboviruses, only Chuzan virus (CHUV) (2.63%) was detected in 2016. The Akabane virus (AKAV), CHUV, Ibaraki virus and Bovine ephemeral fever virus (BEFV) had a positive rate of less than 1.8% in 2017. In antigen test of bovine whole blood, AKAV (12.96%) and BEFV (0.96%) were positive in only one of the farms. As a result of serum neutralization test, antibodies against AKAV were generally measured in all the farms. These results suggest that vaccination before the season in which the Culicoides are active is probably best to prevent arbovirus infections.

Diagnostic value of serum amyloid A in differentiating the inflammatory disorders in wild birds.

Rescued wild birds have very high rates of inflammatory diseases; however, there have been limitations in assessing them sensitively. Few studies have examined acute-phase proteins in wild birds. In this study, serum amyloid A (SAA) was evaluated as an inflammatory indicator along with traditional indices such as white blood cell count, albumin, and albumin/globulin ratio. In total, 291 samples from 139 birds of six avian species were analyzed. All samples were divided into four groups (severe, moderate, mild injuries, and clinically healthy) based on clinical examinations and evaluated by group. SAA levels were measured using an anti-chicken SAA ELISA kit and compared with leukocyte counts, albumin concentrations, and albumin/globulin ratios. Differences among groups were evaluated using the Kruskal-Wallis test, followed by a post hoc test using Dunn's multiple comparisons with SPSS V27. Statistical significance was set at a value of p of <0.05. The median concentration and interquartile range (ng/mL) of SAA in each group were 78.10 ng/mL (50.71-109.30), 31.15 ng/mL (19.85-49.24), 9.68 ng/mL (4.71-22.06), and 3.10 ng/mL (1.50-6.00). We observed a significant difference in the mean levels between the groups (p < 0.001), with the same results observed across species. All four indices showed significant differences in mean values between the groups (p < 0.001). In addition, SAA showed rapid changes in periodically collected samples, reflecting either a positive treatment response or the onset of subclinical diseases. SAA can be used to detect inflammatory conditions and asymptomatic disease in wild birds and is helpful in accurately identifying current health status, which is essential for successful treatment and release.

A preliminary study on effects of storage time and repeated freezing-thawing on the stability of avian serum amyloid A.

Within the field of clinical research, reports on the stability of avian serum amyloid A (SAA) under varying storage conditions are currently scarce. In this study, avian plasma samples were evaluated for SAA, a major acute-phase protein in birds, to assess how varying storage periods and repeated freeze-thaw cycles impact the stability of SAA in the frozen samples. Seven plasma samples from two species and six plasma samples from three species stored at ‒20 °C were used to evaluate the time and temperature effects accordingly. A chicken-specific SAA ELISA kit was used for the measurements. Statistical analysis was performed using SPSS, and the Kruskal-Wallis test and Spearman's correlation coefficient were applied, with statistical significance set at P < 0.05. The SAA concentrations measured daily for 30 days showed no statistically significant differences over time. Freezing-thawing was repeated five times, and a significant negative relationship was confirmed over the cycles (r=‒0.8857, P < 0.05). Although no significance was observed between a decreased concentration and the number of cycles, a decrease in the concentration of > 10% was observed after the fourth cycle in four out of six samples. However, one to three freeze-thaw cycles did not result in a significant decline. Taken together, the results indicate that a negative correlation existed between the mean concentration and multiple freeze-thaw cycles, indicating that these should be avoided where possible.

Evaluation of intrapulmonary arteriovenous anastomoses before and after oxygen supplementation, using transthoracic agitated saline contrast echocardiography in rescued Korean raccoon dogs.

Introduction: Intrapulmonary arteriovenous anastomoses (IPAVAs) are defined as relatively large, dynamic shunt vessels that connect the pulmonary arterial and venous systems, thereby bypassing the pulmonary capillary system. IPAVAs lower elevated pulmonary arterial pressure; however, the presence of the shunt can result in impaired pulmonary gas exchange and paradoxical embolism. Furthermore, the prevalence and effects of IPAVAs in raccoon dogs remain unknown. This study aimed to determine the prevalence of IPAVA among rescued Korean raccoon dogs and evaluate the changes in IPAVA following oxygen supplementation. Methods: Nineteen raccoon dogs rescued by the Jeonbuk Wildlife Centre between August 2022 and December 2023 were subjected to echocardiography. Sixteen healthy and three abnormal raccoon dogs were subjected to transthoracic agitated saline contrast echocardiography (bubble study) based on the echocardiography results. IPAVA was considered to be present if the left heart contrast was visualised after four cardiac cycles following the visualisation of the first right heart contrast. Bubble scores (BS0-5) were assigned based on the maximum number of microbubbles observed in the left ventricular lumen per frame of the ultrasound image. BS was assigned before and after supplementation with 100% oxygen for 5 min. Results: IPAVA was detected in 12 of the 16 healthy raccoon dogs at rest (75%). The BS of the 15 IPAVA-positive raccoon dogs ranged from 1 to 4 points (BS1, 1; BS2, 4; BS3, 8; and BS4, 2). Blood flow through the IPAVA (QIPAVA) was reduced or absent in the 15 IPAVA-positive raccoon dogs after supplementation with 100% oxygen (BS0, 11; BS2, 4). Moreover, BS of the IPAVA showed a significant correlation with the cardiac output per body weight (BW). Conclusion: The prevalence of IPAVA in healthy raccoon dogs at rest was 75%. Adequate oxygen supplementation was found to be effective in reducing QIPAVA, which may help prevent potential negative factors such as pulmonary gas exchange impairments and paradoxical embolism that can occur with IPAVA.

Morbidity of Rescued Wild Birds by Admission Causes in the Republic of Korea.

Insufficient reports are available on what clinical and pathological conditions are observed in rescued free-living wild birds. This study investigated recent diagnoses of admitted wild birds based on admission causes in a southwestern area of South Korea over the past 2 years. A retrospective study was conducted on 1464 birds rescued from 2019 to February 2021. Overall, 12 admission subcategories were classified, and the diagnoses identified for each cause were analyzed. The three most frequently observed categories, general, integumentary, and musculoskeletal, each accounted for 20% of the total diagnoses. Trauma accounted for 71.4% of all diagnoses, and 81.5% featured inflammatory conditions, primarily due to trauma or infection. The proportion of birds that presented inflammatory conditions was much greater than the proportion of birds that were admitted due to trauma-related causes. This was because inflammatory diseases were identified at a high frequency, even from nontraumatic admission causes, and inflammatory conditions were not easily revealed. Suspecting an inflammatory condition in most rescued birds is advisable.

Radiographic and echocardiographic evaluation in rescued Korean raccoon dogs (Nyctereutes procyonoides koreensis).

Introduction: Nyctereutes procyonoides koreensis (Korean raccoon dog), a member of the Canidae family, is anatomically similar to dogs. Previous studies have used vertebral heart scale measurements to measure the cardiac size of Korean raccoon dogs on thoracic radiographs; however, the use of additional cardiac size indices, such as vertebral left arial score, intercostal space, cardiothoracic ratio, and echocardiographic indices, has not been reported. Therefore, this study aimed to establish normal reference ranges for various thoracic radiographic and echocardiographic indices in normal Korean raccoon dogs. Methods: Twenty-six Korean raccoon dogs (11 males and 15 females) were included in this study. The thoracic radiographic indices, vertebral heart scale score, and vertebral left atrial score were measured in the right lateral view. The intercostal space and cardiothoracic ratio were measured in the ventrodorsal view. The echocardiograms were evaluated in the right parasternal long and short axis view and left parasternal apical view. Results: The mean values for the thoracic radiographic and echocardiographic indices were as follows: vertebral heart scale, 9.12 ± 0.74; vertebral left atrial score, 1.5 ± 0.31; intercostal spaces, 3.17 ± 0.34; cardiothoracic ratio, 0.69 ± 0.07; left atrial to aortic root ratio, 1.22 ± 0.14; main pulmonary artery to aorta ratio, 1.22 ± 0.14; left ventricular end-diastolic internal diameter normalized for body weight, 1.36 ± 0.19; end-diastolic volume index, 51.07 ± 19.6; end-systolic volume index, 16.54 ± 7.45; the peak velocity of early diastolic transmitral flow, 73.13 ± 15.46 cm/s; and the ratio between the transmitral flow velocities and the peak early diastolic velocity, 1.77 ± 0.47. Only percent increase in the left ventricular end-systolic internal diameter was negatively correlated with body weight. The remaining indices showed no correlations with body weight. Conclusion: To the best of our knowledge, this is the first case report covering both thoracic radiographic and endocardiographic indices for Korean raccoon dogs. Thus, the thoracic radiographic and echocardiographic indices established in this study may be used to evaluate the cardiac condition of Korean raccoon dogs.

Application of topical rocuronium bromide dosing by ocular size in four species of wild birds.

BACKGROUND: Rocuronium bromide has been evaluated as a mydriatic agent in birds, but the species applied were limited and the dose and effect were variable. OBJECTIVE: This study aims to evaluate the efficacy of topical rocuronium bromide as mydriatics in 4 species according to horizontal palpebral fissure length: Feral pigeon (Columba livia), Common kestrel (Falco tinnunculus), Northern boobook (Ninox japonica), and Eurasian eagle owl (Bubo bubo). METHODS: A total of 32 birds (8 for each species) were included as pre-releasing examination. Rocuronium bromide was instilled in one randomly selected eye of each bird based on palpebral fissure length criteria (0.5 mg/50 µL for pigeons, 1 mg/100 µL for kestrels and boobook owls, and 2 mg/200 µL for eagle owls). The contralateral eye was used as control and treated with normal saline. After instillation of the drug, pupil diameter, pupillary light reflex, intraocular pressure, heart rate, and respiratory rate were evaluated at 10 min intervals up to 180 min and at 30 min intervals up to 360 min. RESULTS: Statistically significant mydriasis was obtained in all birds (p < 0.001). However, in boobook and eagle owls, marked mydriasis persisted until 360 min. Side effects including corneal erosion and lower eyelid paralysis were common, which was observed in 26/32 birds. Blepharospasm was also noted during this study. No systemic adverse signs were observed. CONCLUSIONS: Rocuronium bromide could be a good mydriatics option for 4 species of birds, however, further studies are needed to find lowest effective dose to reduce drug-related side effects.

Antioxidant Effects of Myo-Inositol Improve the Function and Fertility of Cryopreserved Boar Semen.

During cryopreservation, sperm undergoes structural and molecular changes such as ice crystal formation, DNA fragmentation, and reactive oxygen species (ROS) production, leading to decreased sperm quality after thawing. Antioxidants play a crucial role in preventing these damages, both in vivo and in vitro. One potent antioxidant is myo-inositol, known for its protective effects on sperm against ROS. This study aimed to investigate the protective effect of myo-inositol on cryopreserved boar semen. The semen was diluted, cooled, and cryopreserved using a BF5 extender. It was then divided into five groups: control and different concentrations of myo-inositol (0.5, 1, 1.5, and 2 mg/mL). The post-thaw evaluation included assessments of motility, viability, acrosome integrity, mitochondrial membrane potential (MMP), caspase activity, gene expression, ROS levels, apoptosis, and IVF with treated semen. Results showed that myo-inositol at 0.5 mg/mL improved motility, acrosome integrity, and fertilization ability. It also reduced the expression of pro-apoptotic genes and increased SMCP expression. Lower concentrations also demonstrated improved viability and reduced apoptosis and ROS levels. In conclusion, myo-inositol treatment during cryopreservation improved sperm quality, reduced apoptosis and ROS levels, and enhanced fertility rates in boar semen.

Antimicrobial resistance of commensal Escherichia coli and Enterococcus faecalis isolated from clinically healthy captive wild animals in Seoul zoo.

Despite the importance of antimicrobial resistance, only a few studies on the antimicrobial susceptibility on wild animals have been conducted owing to their population, accessibility, and characteristics. The objective of this study was to investigate the prevalence and characteristics of antimicrobial resistance pattern in Escherichia coli and Enterococcus faecalis isolated from the feces of captive wild animals in a zoo. A total of 61 captive wild animals were included in this study. E. coli was isolated from 58 of the 61 animals and E. faecalis was isolated from 29 animals. Among the isolated E. coli strains, ampicillin exhibited the highest resistance rate (27/29, 93.1%). Of these, 18 strains (18/29, 62%) showed multidrug resistance. The multilocus sequence typing (MLST) test showed that only ST155 was detected twice, while the other 16 strains showed different ST types. Among the E. faecalis strains, two were susceptible to all tested antimicrobials, whereas the remaining 27 strains showed resistance to one or more antimicrobials. Nine strains (9/27, 31%) showed multidrug resistance. Among the E. faecalis strains, resistance to quinupristin/dalfopristin was the highest at 96.3% (26/27), while the MLST of the nine MDR strains showed no predominant ST. Genetic association with human isolates or livestock products was observed in the isolated ST types. This indicates that antibiotic resistance in the zoo is responsible for the use of antibiotics and the partial horizontal transmission between humans and animals through feeding or contact.

Case report: Generalized lymphatic anomaly of multiple abdominal organs in a young dog.

A 9-month-old, female Pomeranian dog presented with vomiting and lethargy. Ultrasonography revealed multilobulated anechoic round shape structures at the ovarian and uterine locations. Through computed tomography scan, an extensive non-contrast multilobulated fluid-filled mass suspected of originating from the walls of the ovary, uterus, urinary bladder and rectum was observed. Ovariohysterectomy and urinary bladder biopsy were performed. Histopathological examination revealed numerous cystic lesions lined by plump cuboidal cells believed to be of epithelial origin. Immunohistochemical staining showed that the cyst-like lesions lining cells were strongly positive for lymphatic vessel endothelial hyaluronan receptor 1. Based on these results, lesions were identified as generalized lymphatic anomaly (GLA), in which lymphangiomas develop in multiple organs. After 6 months follow-up, the size of the cysts remaining in the region of the bladder did not undergo much change. GLA should be included in the differential diagnosis when multiple cystic lesions are interspersed in multiple organs.

The presence of anthracosis is associated with the environmental air quality of zoo, wildlife, and companion animals in Jeollabuk-do Province, South Korea.

OBJECTIVE: To determine pulmonary anthracosis in zoo, wildlife, and companion animals of Jeollabuk-do Province, South Korea. ANIMALS: A total of 350 animals of 61 different species, belonging to 3 classes (mammals: n = 38; avian: 21; and reptiles: 2) from different habitats in Jeollabuk-do Province, were examined. PROCEDURES: Gross lung examination and tissue sampling were done at postmortem, and histopathological analysis was microscopically done on hematoxylin and eosin-stained slides. RESULTS: Macroscopic analysis of anthracotic lung tissue revealed minute (pinpoint size) spots and black pigmentation in a scattered and/or coalescing fashion. The presence of carbon particles was noted in 154 (44%, 154/350) cases. Based on habitation, zoo animals had the highest frequency of anthracosis in the lung (55.2%, 69/125), followed by companion animals (45.2%, 56/124) and wildlife animals (28.7%, 29/101). There was an association between habitation and the presence of anthracosis (P < .05). CLINICAL RELEVANCE: This study revealed evidence that the presence of anthracosis is associated with the environmental air quality of zoo, wildlife, and companion animals in Jeollabuk-do Province, South Korea. Air pollution may affect the respiratory health of the endangered species at the Jeonju Zoo as well as the human population. Continuous monitoring of particulate matter and establishing policies that control industrialization around the province would enable quick action to curb any potential respiratory health risks to animals kept in the urban cities of the province.

Altered expression of melanocortin-1 receptor (MC1R) in a yellow-coloured wild raccoon dog (Nyctereutes procyonoides).

BACKGROUND: The melanocortin 1 receptor (MC1R) gene plays a key role in determining coat colour in mammals by controlling the proportion of eumelanin and pheomelanin granules. Wild raccoon dogs have a mixed coat colour, with black to brown and grey hairs. HYPOTHESIS/OBJECTIVES: The study was performed to identify the cause of the variant yellow coat colour in a wild raccoon dog. ANIMALS: A wild raccoon dog that showed coat colour change to yellow and four wild-type raccoon dogs that showed normal coat colour were included. METHODS: To identify the cause of the variant yellow coat colour, we examined the sequence of the MC1R gene and its expression at the mRNA and protein levels. RESULTS: The coding region of the MC1R gene of this raccoon dog comprised 954 bp, the same as for wild-type raccoon dogs and domestic dogs. By comparing the gene with that in the wild-type raccoon dog, a 2 bp deletion was detected in the 5'-untranslated region, positioned 152 bp upstream of the start codon. However, there was no significant difference in the mRNA expression level. The yellow raccoon dog revealed a significantly decreased MC1R protein level compared with the wild-type raccoon dogs, indicating an increase in pheomelanin synthesis. CONCLUSIONS AND CLINICAL IMPORTANCE: These results suggest that the variant coat colour in the yellow raccoon dog was associated with decreased MC1R function.

Isolation of Staphylococcus simulans from dermatitis in a captive African pygmy hedgehog.

Staphylococcus simulans, a coagulase-negative staphylococcal species, can cause several diseases in humans and animals. This report describes a case of dermatosis characterized by broad, well-circumscribed hyperkeratosis and alopecia on the back of a household pygmy hedgehog (Erinaceous albiventris). Quills and exudates were studied by microscopy. The microscopic examination of the exudates collected from the lesion revealed several leukocytes and numerous gram-positive cocci. An aerobic bacterial culture revealed overgrowth of the same gram-positive bacteria. The bacterium was identified as S. simulans by polymerase chain reaction amplification and direct sequencing targeted to the 16S ribosomal RNA gene. This report is the first to show that S. simulans could be related to the dermatitis of hedgehogs.

Distribution and epidemiological relatedness of methicillin-resistant Staphylococcus aureus isolated from companion dogs, owners, and environments.

This study aimed to investigate the distribution and epidemiological relatedness of methicillin-resistant Staphylococcus aureus (MRSA) isolates from companion dogs, owners, and residential environments of 72 households. Sampling was performed twice from January to June 2018 and a total of 2,592 specimens were collected. The specimens collected from each household were streaked on CHROM agar S. aureus and the colonies grown on the medium were further identified using a mass spectrometry microbial identification system. Antimicrobial susceptibility testing, Panton-Valentine-Leukocidin (PVL) gene PCR, staphylococcal cassette chromosome mec (SCCmec) typing, Staphylococcus aureus Protein A (spa) typing, pulsed-field gel electrophoresis (PFGE), and multi-locus sequence typing (MLST) were conducted to evaluate the phenotypic and genotypic characteristics of the MRSA isolates. A total of 65 S. aureus strains (2.5%) were isolated and 49 (1.9%) of 65 strains were MRSA displaying cefoxitin-resistance with mecA carriage. MRSA strains were isolated from dogs (n=6, 9.2%), owners (n=27, 41.5%), and residential environments (n=16, 24.6%), respectively. Overall prevalence of non-duplicated MRSA was 16.7% (12/72 households) at household level. ST72-SCCmec IVc MRSA clones predominantly appeared in MRSA-positive families. Furthermore, PFGE analyses showed that ST72-SCCmec IVc-t324 is shared between dog owners and dogs. To our knowledge, this is the first study to report the sharing of ST72 MRSA between dogs and their owners.

Identification of Zoonotic Tick-Borne Pathogens from Korean Water Deer (Hydropotes inermis argyropus).

Korean water deer (Hydropotes inermis argyropus) are widespread in the Republic of Korea (ROK). Mostly, Korean water deer are essential hosts for maintaining ticks and tick-borne diseases (TBDs). Here, we investigated the prevalence of tick-borne pathogens (TBPs) among rescued Korean water deer. Anaplasma phagocytophilum (21.4%, 6/28), Anaplasma capra (14.3%, 4/28), Babesia capreoli (3.6%, 1/28), and Coxiella burnetii (3.6%, 1/28) were detected, but Borrelia burgdorferi, Ehrlichia, Rickettsia, and Theileria infections were not found. A. phagocytophilum was the most commonly detected pathogen, and co-infection with A. capra and B. capreoli was also noted in one Korean water deer. To our knowledge, this is the first article of B. capreoli infection in Korean water deer in the ROK. The infecting isolate of A. phagocytophilum was genetically characterized by 16S ribosomal RNA (rRNA) gene and ankyrin-related protein (ankA) gene. Although the 16S rRNA gene alone may not be informative enough to delineate distinct host species, ankA-based phylogeny revealed a high identity of Korean water deer sequences with those of the causative agent of human granulocytic anaplasmosis. A. capra was detected by using citrate synthase gene (gltA), heat-shock protein (groEL), and major surface protein 4 (msp4) genes. Phylogenetic tree based on these gene markers revealed that there were at least two distinct variants within A. capra circulating in the ROK. One variant originated from different hosts including humans, ticks, goats, and sheep, whereas the other variant was reported recently in Korean water deer in the ROK. Consequently, these sequences were identified to belong to a zoonotic species. Sequencing analysis of the 18S rRNA gene revealed that our isolate belonged to B. capreoli and was distinct from Babesia divergens and Babesia venatorum. Moreover, our isolate showed 92.2% homology with B. capreoli sequences, indicating that these differences may be attributed to the different tick species that transmit B. capreoli or to different host species. Genotyping and phylogenetic analysis of C. burnetii based on 16S rRNA and IS1111 genes revealed that our isolate was grouped with several strains of C. burnetii and was genetically distant from Coxiella-like bacteria isolates. The present results highlight that Korean water deer act as potential reservoir hosts for zoonotic TBPs, and thus play an important role in the transmission of TBDs in humans, animals, and livestock.

Prevalence and genotypes of Enterocytozoon bieneusi in wildlife in Korea: a public health concern.

BACKGROUND: Enterocytozoon bieneusi is a unicellular microsporidian fungal pathogen that infects a broad range of animal hosts, including wild and domestic animals and humans. The infection burden of this parasite in wild animals in Korea is largely unknown. In this study, the occurrence and genotypes of E. bieneusi were investigated in wild animal populations in Korea. METHODS: A total of 157 fecal samples (97 from Korean water deer, 48 from raccoon dogs and 12 from other taxa) were collected from wild animals at five wildlife centers in Korea. Genomic DNA was extracted from the samples and screened by nested-PCR targeting the internal transcribed spacer (ITS) region of rRNA, followed by sequence analysis to determine the genotype(s) of E. bieneusi. RESULTS: The overall prevalence of E. bieneusi was 45.2% (71/157), with rates of 53.6% (52/97) in Korean water deer, 35.4% (17/48) in raccoon dogs and 16.7% (2/12) in other taxa. We detected seven ITS genotypes, including one known (genotype D) and six new genotypes (Korea-WL1-Korea-WL6). Phylogenetically, all detected genotypes clustered with counterparts belonging to group 1, which includes isolates from different animal hosts and humans, suggesting their zoonotic potential. CONCLUSIONS: Our survey results indicate that E. bieneusi circulates widely in wild animals in Korea. These findings address the role of wildlife as a potential source of microsporidiosis in domestic animals and humans.