Young Kyu Kim

Behavioural reactivity of the Korean native Jindo dog varies with coat colour.

This study aimed to compare the behavioural reactivity of Jindo dogs with two different coat colours. Fawn (16 males, 15 females; mean age+/-S.D.=7.2+/-2.1 years) and white (10 males, 10 females; mean age+/-S.D.=6.9+/-2.1 years) Jindo dogs were exposed to a set of behavioural tests. All of the dogs were videotaped during the testing period to allow further analysis. The intensity of social, aggressive, fearful, and submissive reactivity and the frequency of urination as a scent-marking behaviour were scored on a scale running from 0 to 4 points. For each dog, each variable was defined as the average of the scores of nine behaviour tests. Then, the behavioural reactivities of Jindo dogs of each coat colour were compared. The results suggested that Jindo dogs of fawn coat colour exhibited a significantly lower intensity of fearful and submissive reactivity than those of white coat colour. In addition, fawn Jindo dogs produced scent-marking behaviour significantly more frequently. The results of the present study may provide useful information for scientific researchers, potential owners and breeders of Jindo dogs.

Sprayed intraperitoneal bupivacaine reduces early postoperative pain behavior and biochemical stress response after laparoscopic ovariohysterectomy in dogs.

This study investigated the use of sprayed intraperitoneal bupivacaine to relieve postoperative pain behavior and biochemical stress response after laparoscopic ovariohysterectomy (LOVH) in dogs. Sixteen sexually intact female dogs were randomly assigned to two groups. The sprayed intraperitoneal bupivacaine (SIB) group received 4.4 mg/kg of sprayed intraperitoneal bupivacaine diluted to 0.25% with an equivalent volume of saline after pneumoperitoneum. The control group received 1.76 mL/kg of saline in a similar fashion. Both groups received preoperative periportal 5% bupivacaine (1 mL) before incision. Postoperative pain was measured using the short form of the Glasgow composite measures pain scale (CMPS-SF, 0-24). Serum cortisol and glucose concentrations were measured preoperatively and 0.5, 1, 2, 4, 6, 12, and 24h postoperatively. The SIB group had significantly lower CMPS-SF compared to the control group 1, 2, 4, 6, and 12h after the operation. Cortisol concentrations were significantly increased from preoperative concentrations in the control group at 0.5, 1, 2, and 4h post operation and at 0.5 and 1h post operation in the SIB group. No significant differences were seen in serum glucose within each group. This report suggests that the use of sprayed intraperitoneal bupivacaine can be used as part of a multimodal approach for pain management after LOVH in dogs.

RNA-seq-based metatranscriptomic and microscopic investigation reveals novel metalloproteases of Neobodo sp. as potential virulence factors for soft tunic syndrome in Halocynthia roretzi.

Bodonids and trypanosomatids are derived from a common ancestor with the bodonids being a more primitive lineage. The Neobodonida, one of the three clades of bodonids, can be free-living, commensal or parasitic. Despite the ecological and evolutionary significance of these organisms, however, many of their biological and pathological features are currently unknown. Here, we employed metatranscriptomics using RNA-seq technology combined with field-emission microscopy to reveal the virulence factors of a recently described genus of Neobodonida that is considered to be responsible for ascidian soft tunic syndrome (AsSTS), but whose pathogenesis is unclear. Our microscopic observation of infected tunic tissues suggested putative virulence factors, enabling us to extract novel candidate transcripts; these included cysteine proteases of the families C1 and C2, serine proteases of S51 and S9 families, and metalloproteases grouped into families M1, M3, M8, M14, M16, M17, M24, M41, and M49. Protease activity/inhibition assays and the estimation of expression levels within gene clusters allowed us to identify metalloprotease-like enzymes as potential virulence attributes for AsSTS. Furthermore, a multimarker-based phylogenetic analysis using 1,184 concatenated amino acid sequences clarified the order Neobodo sp. In sum, we herein used metatranscriptomics to elucidate the in situ expression profiles of uncharacterized putative transcripts of Neobodo sp., combined these results with microscopic observation to select candidate genes relevant to pathogenesis, and used empirical screening to define important virulence factors.

Heat shock protein profiles on the protein and gene expression levels in olive flounder kidney infected with Streptococcus parauberis

Heat shock proteins (HSPs) have been observed in cells exposed to a variety of stresses, including infectious pathogens. This study used a label-free, quantitative proteomic approach and transcriptional gene expression analysis to investigate infection-related HSP proteins and their encoding genes in whole kidneys from olive flounder (Paralichthys olivaceus). During Streptococcus parauberis infection in the flounder, the genes encoding Hsp10, Hsp40A4, Hsp40B6, Hsp40B11, Hsp60, Hsp70, glucose regulated protein 78 (Grp78), Hsp90α, Hsp90β and Grp94 were induced, and the protein levels of Hsp60, Hsp70, Hsp90α, Hsp90β and Grp94 were differentially regulated over time. Subsequent results also revealed that Hsp60, Hsp70, Hsp90α, Hsp90β and Grp94 appear to be the dominant and critical HSPs in olive flounder during bacterial infection. This is the first estimation of the differential involvement of HSPs in the immune response of olive flounder exposed to bacterial infection.

LGP2 Expression is Enhanced by Interferon Regulatory Factor 3 in Olive Flounder, Paralichthys olivaceus

In innate immunity, LGP2 (laboratory of genetics and physiology 2) plays a very important role in the production of type I interferon (IFN) through recognition of cytosolic viral RNA. Although viral infection or stimulation with double-strand RNA dramatically induces expression of the LGP2 gene, the underlying transcriptional mechanism has never been studied. Here, we cloned and characterized the 5′-upstream region (−1,337 bp) of the LGP2 gene in olive flounder (Paralichthys olivaceus). Numerous canonical motifs for IFN-regulatory factors (IRFs) were found in this region, and reporter assays identified a poly I:C-responsive promoter region (−506 to −398) that regulated LGP2 transcription. Transcriptional activity of the LGP2 promoter was strongly enhanced by IRF3, which bound to IRF3 motif #3 (−480). The LGP2 promoter was also responsive to viral infection in vitro. These results suggest that LGP2 transcriptional control is crucially involved to regulated by IRF3 function after viral infection or stimulation with poly I:C.

Development of a multiplex PCR assay to detect Edwardsiella tarda, Streptococcus parauberis, and Streptococcus iniae in olive flounder (Paralichthys olivaceus)

A multiplex PCR protocol was established to simultaneously detect major bacterial pathogens in olive flounder (Paralichthys olivaceus) including Edwardsiella (E.) tarda, Streptococcus (S.) parauberis, and S. iniae. The PCR assay was able to detect 0.01 ng of E. tarda, 0.1 ng of S. parauberis, and 1 ng of S. iniae genomic DNA. Furthermore, this technique was found to have high specificity when tested with related bacterial species. This method represents a cheaper, faster, and reliable alternative for identifying major bacterial pathogens in olive flounder, the most important farmed fish in Korea.

Laparoscopic nephrectomy in dogs: An initial experience of 16 experimental procedures

Left laparoscopic nephrectomy was performed in 16 dogs to describe the surgical techniques and initial experiences associated with operation time and surgery complications. The renal vein and artery were occluded by three ligating clips, respectively, and the ureter was sectioned after ligation with ligating clips at the level of the iliac vessels. A morcellation technique was used to remove the kidney from the abdominal cavity after placing it into a specimen retrieval bag. Total operation time and time spent for each different surgical stage in the first five operations were compared with those in the last five of the 16 operations. The factors that affected the differences of total operation time were examined, including sex, bodyweight, number of operations, incision length, and surgical stages. Six intra-operative complications occurred including splenic hemorrhage (3 cases), torn specimen retrieval bag during kidney morcellation (1 case), and subcutaneous emphysema (2 cases). Surgical time for laparoscopic nephrectomy was affected primarily by the time spent for renal vascular pedicle section and could be decreased as the number of cases increased. Thus, laparoscopic nephrectomy using ligating clips and morcellation for kidney removal could be considered where nephrectomy is indicated in dogs.

Characterization of a specific monoclonal antibody against immunoglobulin light kappa/L1 chain in olive flounder (Paralichthys olivaceus)

ABSTRACT Immunoglobulins (Ig) are heterodimeric proteins that play critical roles in the adaptive immune system of vertebrates. Because of their plasticity, teleostean Igs are more diverse, and thus do not conform to mammalian classifications. Because of this, mammalian‐based Ig cell markers cannot be used successfully to study immune responses in fish. There is therefore a need to produce Ig‐specific cell markers for fish. Here, we attempted to identify the specific isotype detected by an Ig light chain‐specific monoclonal antibody (anti‐olive flounder IgL‐mAb: M7C3–4) that we had previously produced [11]. Three newly identified sequences of the Ig light chain from olive flounder were classified according to their isotypes. Subsequent analyses revealed that M7C3–4 was able to specifically detect lymphocytes expressing one of the &kgr; chains (Ig&kgr;‐a) in olive flounder. Interestingly, Ig&kgr;‐a+ B cells were more abundant in spleen and trunk‐kidney than in peripheral blood, indicating a distribution different from that of IgM+ B cells. Our work reveals interesting aspects of B cell distribution and differentiation, and may aid in the production of suitable and effective cell markers for olive flounder. HIGHLIGHTSOlive flounder has three isotypes of IgL, two of &kgr;, and one of &sgr;.M7C3–4 produced against serum immunoglobulins of olive flounder specifically recognizes Ig&kgr;‐a isotype.Ig&kgr;‐a+ B cells are more abundant in organs than in peripheral blood.