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.

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.

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.

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.

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.

Systemic Infection of Mycobacterium abscessus in a Free-Ranging Wild Eurasian Eagle Owl (Bubo bubo).

An adult Eurasian Eagle Owl (Bubo bubo) rescued from drowning was unable to fly. After euthanasia, necropsy and histopathologic examination showed granulomatous inflammation and intracellular acid-fast stain-positive rod-shaped bacteria in the skin, lung, liver, and spleen, which were identified by using molecular analysis as Mycobacterium abscessus.

Alu-derived old world monkeys exonization event and experimental validation of the LEPR gene.

The leptin receptor (LEPR) is a crucial regulatory protein that interacts with Leptin. In our analysis of LEPR, novel AluJb-derived alternative transcripts were identified in the genome of the rhesus monkey. In order to investigate the occurrence of AluJb-derived alternative transcripts and the mechanism underlying exonization events, we conducted analyses using a number of primate genomic DNAs and adipose RNAs of tissue and primary cells derived from the crab-eating monkey. Our results demonstrate that the AluJb element has been integrated into our common ancestor genome prior to the divergence of simians and prosimians. The lineage-specific exonization event of the LEPR gene in chimpanzees, orangutans, and Old World monkeys appear to have been accomplished via transition mutations of the 5' splicing site (second position of C to T). However, in New World monkeys and prosimians, the AluJb-related LEPR transcript should be silenced by the additional transversion mutation (fourth position of T to G). The AluJb-related transcript of human LEPR should also be silenced by a mutation of the 5' splicing site (first position of G to A) and the insertion of one nucleotide sequence (minus fourth position of A). Our data suggests that lineage-specific exonization events should be determined by the combination event of the formation of splicing sites and protection against site-specific mutation pressures. These evolutionary mechanisms could be major sources for primate diversification.

Processing and subcellular localization of ADAM2 in the Macaca fascicularis testis and sperm.

Fertilin, a heterodimeric protein complex composed of ADAM1 and ADAM2 located on the sperm surface, is involved in sperm-egg interaction. In our study, we examined the physiological processing and subcellular localization of M. fascicularis ADAM2 during spermatogenesis in the testis and epididymal tract. M. fascicularis ADAM2 was initially synthesized as a 100 kDa precursor in testicular germ cells. After passing into 50 kDa intermediate form in the epididymal tracts, the precursor form was finally processed into a 47 kDa protein in sperm. We found that M. fascicularis ADAM2 is localized on the sperm surface and contributes to the formation of a candidate fertilin complex. In particular, Far-Western blot analysis revealed that M. fascicularis ADAM2 cystein-rich domain may be related to protein-protein interaction. Therefore, the cystein-rich domain of ADAM2 could provide a mechanism to form a fertilin complex.

Structural modifications of outer membrane vesicles to refine them as vaccine delivery vehicles.

In an effort to devise a safer and more effective vaccine delivery system, outer membrane vesicles (OMVs) were engineered to have properties of intrinsically low endotoxicity sufficient for the delivery of foreign antigens. Our strategy involved mutational inactivation of the MsbB (LpxM) lipid A acyltransferase to generate OMVs of reduced endotoxicity from Escherichia coli (E. coli) O157:H7. The chromosomal tagging of a foreign FLAG epitope within an OmpA-fused protein was exploited to localize the FLAG epitope in the OMVs produced by the E. coli mutant having the defined msbB and the ompA::FLAG mutations. It was confirmed that the desired fusion protein (OmpA::FLAG) was expressed and destined to the outer membrane (OM) of the E. coli mutant from which the OMVs carrying OmpA::FLAG are released during growth. A luminal localization of the FLAG epitope within the OMVs was inferred from its differential immunoprecipitation and resistance to proteolytic degradation. Thus, by using genetic engineering-based approaches, the native OMVs were modified to have both intrinsically low endotoxicity and a foreign epitope tag to establish a platform technology for development of multifunctional vaccine delivery vehicles.

Importance of the porcine ADAM3 disintegrin domain in sperm-egg interaction.

In the mouse, ADAM3, a well-characterized testis-specific protein of the A disintegrin and metalloprotease (ADAM) family, has a crucial role in fertilization by mediating sperm binding to the egg zona pellucida. However, little is known about ADAM3 in other species, such as domestic pigs. We have identified porcine ADAM3 and analyzed the protein. RT-PCR and trypsinization of sperm surface proteins revealed that porcine ADAM3 is expressed at high levels in the testis and on the sperm surface. Furthermore, an IVF inhibition assay with a recombinant porcine ADAM3 disintegrin domain showed that treatment of the disintegrin domain effectively prevented pig sperm-egg interactions. In the present study, we demonstrated the presence of ADAM3a and ADAM3b molecules in the pig and examined their roles in fertilization.

Identification of novel retromer complexes in the mouse testis.

A family of vacuolar protein sorting (Vps) proteins, which are components of mammalian retromer complex, has been studied in the mouse. Vps26a is known as a retromer component that plays an important role in embryonic development: however, its cell-type expression and precise role remain to be elucidated. In this study, we identified a new isoform of Vps26a, called Vps26aT, which was expressed specifically in the mouse testis. Diverse expression patterns of Vps26 variants in mouse tissues were determined by Western blot and RT-PCR analyses, and the direct interaction of Vps26aT with Vps35 was also demonstrated by immunoprecipitation and pull-down assay using antibodies raised against each Vps component. Our results revealed that the retromer complex could be formed from different Vps26 isoforms in a tissue-specific manner, resulting in more than two types of the retromer complex, including the Vps26a-Vps29-Vps35, Vps26aT-Vps29-Vps35, and Vps26b-Vps29-Vps35 complexes in mouse tissues.

Use of microsatellite markers to detect quantitative trait loci in Yorkshire pigs.

To identify genetic markers associated with economic traits in pigs, 157 microsatellite markers were examined in Yorkshire pigs. Thirty eight female Yorkshire pigs were initially examined and six of them were selected as progenitors; half were more than 1.5 standard deviations (SD) above the mean for average daily gain (ADG) and backfat thickness (BFT), and the remaining half were more than 1.5 SD below the mean. These pigs were then mated to male Duroc pigs, and 200 F2 pig offspring were examined for the association of specific alleles with ADG and BFT. To confirm the specific markers identified in the initial analysis, associations of significant markers with economic traits were further examined in 228 additional performance-tested purebred pigs. Twenty-five microsatellite markers were significantly associated with either ADG or BFT, and among these, 17 were associated with both traits. The markers with the highest association to ADG were also associated with BFT. Our study reveals that specific markers could be used to predict economic significance, and confirms several quantitative trait loci (QTL) identified in previous studies. However, further analysis with more closely-spaced microsatellite markers is required to refine predictive values for economic traits and positions of QTL that are reliable for actual phenotypic prediction.