The lateral cytoplasmic processes of tenocytes extend to form three-dimensional network surrounding collagen fibers. It is unknown whether connections between two cytoplasmic processes involve overlapping of the processes or merely surface contact. In this study, the two-dimensional and three-dimensional structure of tenocytes in the Achilles tendons of the newly hatched chicks were studied using transmission electron microscopy and serial block face-scanning electron microscopy. Observation of the two-dimensional structures revealed various forms of cellular connections, including connections between the cytoplasmic processes of adjacent tenocytes and between the cytoplasmic process of tenocytes and fibroblasts. Three-dimensional observation showed spike-like cytoplasmic processes extending from one tenocyte that interlocked with cytoplasmic processes from other tenocytes. Cytoplasmic processes from each tenocyte within the chick tendons interlocked to ensure a tight cell-to-cell connection around growing collagen fibers. A cellular network formed by these cytoplasmic processes surrounds each collagen fiber. Cell-cell junctions, which were suggested to be gap junctions, observed at sites of cytoplasmic process overlap most likely represent the major route for communication between tenocytes associated with fibroblasts, enabling vital signals important for maintaining the cell and tendon integrity to be transmitted.

The comprehensive targets of innervation in the intestinal mucosa are unknown, partly because of the diversity of cell types and the complexity of the neural circuits. Herein, we investigated the comprehensive targets of neural connectivity and analyzed the precise characteristics of their contact structures in the mucosa of rat ileum. We examined target cells of neural connections and the characteristics of their contact structures by serial block-face scanning electron microscopy at four portions of the rat ileal mucosa: the apical and basal portions in the villi, and the lateral and basal portions around/in the crypts. Nerve fibers were in contact with several types of fibroblast-like cells (FBLCs), macrophage-like cells, eosinophils, lymphocyte-like cells, and other types of cells. The nerve fibers almost always ran more inside of lamina propria than subepithelial FBLC, and thus contacts with epithelial cells were very scarce. The contact structures of the nerve fibers were usually contained synaptic vesicle-like structures, and we classified them into patterns based on the number of nerve fiber contacting the target cells at one site, the maximum diameter of the contact structures, and the relationship between nerve fibers and nerve bundles. The contact structures for each type of cells occasionally dug into the cellular bodies of the target cells. We revealed the comprehensive targets of neural connectivity based on the characteristics of contact structures, and identified FBLCs, immunocompetent cells, and eosinophils as the candidate targets for innervation in the rat ileal mucosa.

1-kestose is a structural component of fructo-oligosaccharides and is composed of 2 fructose residues bound to sucrose through β2-1 bonds. In the present study, the influence of the ingestion of 1-kestose on the intestinal microbiota was investigated in cats. Six healthy cats were administered 1 g/day of 1-kestose for 8 weeks followed by a 2-week wash-out period. Fecal samples were collected from cats after 0, 4, 8, and 10 weeks. The intestinal microbiota was examined by a 16S rRNA gene metagenomic analysis and real-time PCR. Short-chain fatty acids were measured by GC/MS. The results suggested that the intestinal bacterial community structure in feline assigned to this study was divided into 2 types: one group mainly composed of the genus Lactobacillus (GA) and the other mainly composed of the genus Blautia with very few bacteria of Lactobacillus (GB). Furthermore, the number of Bifidobacterium slightly increased after the administration of 1-kestose (at 4 and 8 weeks) (P<0.1). The administration of 1-kestose also increased the abundance of Megasphaera, the butyric acid-producing bacteria, at 4 and 8 weeks (P<0.1). Furthermore, an increase in butyric acid levels was observed after the administration of 1-kestose for 4 weeks (P<0.1). These results suggest that 1-kestose activated butyrate-producing bacteria as well as bifidobacteria and propose its potential as a new generation prebiotic.

A 12-year-old female domestic short-haired cat was presented due to weight loss, anorexia, and tachypnea. Complete blood count revealed severe anemia, leukocytosis with massive undifferentiated blast cells, and thrombocytopenia. Bone marrow aspiration showed acute myeloid leukemia, subclassified as monoblastic leukemia (M5a) based on the outcomes of the cytochemistry examinations. The SNAP feline leukemia virus (FeLV) and feline immunodeficiency virus (FIV) test using whole blood was negative. In addition, FeLV/FIV proviral polymerase chain reaction test using bone marrow aspirate was also negative. Although the cat was treated with doxorubicin, cytosine arabinoside, and prednisolone, anemia did not improve without blood transfusion. The owner declined further treatment after 2 months, and the cat died a few days later.

Polymerase chain reaction (PCR) is typically used for the early detection of mycoplasma in bovine milk; it requires 3 days to obtain results because of the necessary enrichment process. A more rapid, simple, and accurate detection method is required to directly detect the Mycoplasma bovis (M. bovis) gene in milk. In this study, we assess the utility of combining the following two methods to achieve this goal: the loop-mediated isothermal amplification (LAMP), which is more sensitive than PCR, and the procedure for ultra rapid extraction (PURE), which adsorbs and filters components that inhibit DNA amplification/detection. LAMP was examined using DNA extracts obtained by four methods. This showed that PURE had the highest sensitivity and specificity and that the combination of PURE and LAMP was able to detect M. bovis in milk. We then showed that the detection limit of M. bovis was 10² colony-forming units per milliliter of milk using the PURE–LAMP. Finally, the respective sensitivities of the PURE–LAMP and PCR were 57% and 86% for bulk tank milk, 89% and 74% for mature milk, 85% and 92% for colostrum/transitional milk, and 97% and 95% for mastitis milk. The specificity was 100% for all milk samples in both LAMP and PCR. We conclude that PCR was suitable for detecting mycoplasma in bulk tank milk and that the PURE–LAMP could detect mycoplasma within 2 hr and was also effective for mature and mastitis milk.

We investigated changes in oxidative stress markers during the transition period in healthy Holstein cows and those with postpartum diseases. Transition control (TC) Holstein cows (n=9) were evaluated for longitudinal changes during the transition period and postpartum diseased (PD) cows with ketosis (n=10), abomasal displacement (n=9), and acute mastitis (n=10) were evaluated in comparison to control cows (n=10). In the TC group, blood samples were collected at 2 weeks prepartum and at 1, 2, 4, 6, and 8 weeks postpartum. Milk yield and composition were measured at 2 and 4 weeks postpartum. In the PD group, blood samples were collected at the first day of examination during the 60 days postpartum. Peripheral oxidative stress parameters (malondialdehyde, MDA; potential antioxidant capacity, PAO; and glutathione peroxidase) were measured, and biochemical analyses were performed. In the TC group, MDA increased significantly postpartum and was correlated with milk yield, blood glucose (Glu), free fatty acid (FFA), β-hydroxybutyric acid (BHB), and aspartate aminotransferase. Compared to the control cows, PD cows with ketosis had significantly higher MDA and significantly lower PAO. Moreover, MDA was significantly correlated with Glu, FFA, and BHB. Postpartum increase in MDA might interact with milk yield and Glu, FFA, and BHB in the TC cows, and postpartum diseases, especially ketosis, might signify its increase and interaction with Glu, FFA, and BHB.

The objective of the present study was to elucidate sequential changes in mRNA abundance of serum amyloid A (SAA) isotypes in endotoxin (ETX) challenge model cattle. Ten healthy cattle were separated to 2 groups: control and ETX groups. Cattle in the ETX group were challenged by 2.5 µg/kg of O111:B4 lipopolysaccharide in 4 ml of autologous serum. Blood samples were withdrawn at pre, 0.5, 1, 2, 4, 8, 12, 24, 48, 72 and 96 hr after ETX challenge. Plasma ETX activity, serum SAA concentrations, mRNA abundance of interleukin (IL)-6, SAA2 and SAA4 in the liver and polymorphonuclear leukocytes were measured. The plasma ETX activity in the ETX group increased at 0.5 hr after the ETX challenge. The serum SAA value remained higher between 12 and 72 hr after the ETX challenge than that of the control group. Hepatic IL-6 mRNA abundance in the ETX group increased at 2 hr after the ETX challenge. Hepatic SAA2 and SAA4 mRNA abundance significantly increased from 4 hr after administration, and remained significantly higher than those pre-values up to 12 and 24 hr, respectively. The abundance ratio of hepatic SAA2 was much higher than that of SAA4. The major isotype was SAA2 in liver tissue, and it is indicating systemic inflammation in cattle.

The feasibility of ultrasonographic measurement of thyroid gland area to common carotid artery (TG:CCA) was investigated. Twenty-one healthy, 12 hypothyroid and 18 non-thyroid illness (NTI) dogs were evaluated. The area of thyroid lobe and common carotid artery in right and left sides were measured using the same ultrasonographic images in transverse plane. The average of the right and left ratio was calculated as TG:CCA. The median TG:CCA of 21 healthy dogs was 1.53, and it did not correlate either body weight or age. The median TG:CCA of 12 hypothyroid dogs was 0.81, which was significantly lower than that of 18 NTI dogs (1.81, P<0.001). If the cut off value <1.12 was used, TG:CCA indicated hypothyroidism with a sensitivity of 100%, specificity of 83%, and accuracy of 90%. Our data indicated that TG:CCA was independent of both body weight, which may contribute to consistent measurement of thyroid size. The results of this study suggest that TG:CCA is a promising tool for diagnosing canine hypothyroidism.

This study was performed to confirm the alterations of blood and urine parameters in artificially induced hypocalcemic cows. For a 2 × 2 cross-over design, four non-pregnant, non-lactating Holstein Friesian cows (623 ± 63 kg) were utilized. Cows in the treatment and control group were infused with ethylenediaminetetraacetic acid (Na₂EDTA) solution and normal saline through an intravenous catheter for 3 hr, respectively. Laboratory analyses included complete blood cell count, plasma chemistry, blood gas analysis and urine chemistry. During the hypocalcemic period, abnormal signs were not observed clinically, hematologically nor biochemically either in groups. But, plasma calcium and magnesium concentrations continued to decrease throughout Na₂EDTA infusion, and significant group differences (P<0.05 or P<0.001) were detected until 5 hr after the initiation of infusion. Urinary excretions of these minerals were significantly reduced compared to the control group by 6 hr (Ca, P<0.05; Mg, P<0.001). Moreover, there is a significant group difference in the change in plasma pH at 1 hr after Na₂EDTA infusion (P<0.05) and maintained a decreased level until 6 hr. Consequently, the blood pH was diminished simultaneously with hypocalcemia and hypomagnesemia induction in cows infused with Na₂EDTA. This phenomenon may be one of the mechanisms to recover normocalcemia including maximizing the effect of parathyroid hormone, however, further studies are needed to elucidate the mechanism to alter the blood pH in hypocalcemia.

To investigate the prevalence of murine astrovirus (MuAstV) in mice in laboratory animal facilities in Japan, a polymerase chain reaction (PCR) test targeting the RNA-dependent RNA polymerase (RdRP) gene was performed on the cecum contents of 1,212 mice (1,183 immunocompetent mice and 29 immunodeficient mice) from 226 facilities. The results showed that 118 (52.2%) of the 226 facilities were positive for MuAstV. Out of the 1,212 mice, 424 (35.0%) were positive. No gross lesions were observed in any of the mice examined. A phylogenetic analysis for 15 selected strains revealed that 13 strains formed one cluster, while two were genetically distant from that cluster. These results suggest that multiple strains are prevalent in laboratory mice in Japan.

Tick-borne diseases (TBD) cause enormous losses for farmers. Backyard raising comprises majority of the livestock population in the Philippines, but TBD information in backyard livestock is scarce. In this study, 48 cattle and 114 water buffalo samples from Quezon province, Philippines were molecularly screened for tick-borne pathogens. Anaplasma marginale (16.67%) and hemoplasma (20.99%) were detected in the samples. A. marginale infection (P=0.0001) was significantly higher in cattle, while hemoplasma infection (P=0.011) was significantly higher in water buffaloes. A. marginale isolates from this study were highly similar to previous isolates from the Philippines while Mycoplasma wenyonii and Candidatus Mycoplasma haemobos were the identified hemoplasma species. Our findings reveal additional information on the TBD situation of Philippine backyard livestock.

Interleukin (IL)-19 is a cytokine of the IL-10 family. There are many reports on the involvement of IL-19 in several human diseases. There also are many reports elucidating the role of IL-19 using mouse disease models. Most reports use C57BL/6 mice, whereas few reports use BALB/c mice, in terms of the mouse disease model that the researchers used in the present study. To date, research on the role of IL-19 is diversified, yet some basic mechanisms are still unclear. In this study, we administered lipopolysaccharide (LPS), polyI:C, and CpG to BALB/c mice, measured more than 20 cytokines in the blood and compared them with that of the wild-type and IL-19-deficient (IL-19 KO) mice. LPS is associated with bacterial infection, polyI:C is associated with viral infection, and CpG is associated with both bacterial and viral infections. Among the cytokines measured, the results of experiments using LPS revealed that the production of some cytokines was suppressed in IL-19 KO mice. Interestingly, the experiments using polyI:C revealed that production of some cytokines was enhanced in IL-19 KO mice. However, the experiments using CpG have shown that the production of only one cytokine was enhanced in IL-19 KO mice. These results revealed that cytokine production in the blood was regulated by IL-19, and the type of regulation was dependent on the administered stimulant.

Extracellular vesicles (EV) consist of a lipid-bilayered membrane and are typically classified as small EV (sEV or exosome) or large EV (or microvesicle). sEV mediate cell-to-cell communication and play a key role in various disease states. We recently reported that plasma sEV in normotensive Wistar Kyoto rats (WKY) and spontaneously hypertensive rats (SHR), an animal model of human essential hypertension, regulate systemic blood pressure (BP). An abnormal vascular reactivity is involved in the onset and progression of hypertension. In the present study, we tested the hypothesis that plasma sEV may affect the reactivity of isolated blood vessels. sEV were isolated from plasma in male WKY and SHR (WsEV and SsEV, respectively) by precipitation with polyethylene-glycol and ultracentrifugation. The particle distribution and concentration of sEV were measured by a tunable resistive pulse sensing method. Isolated mesenteric arteries from normal male Wistar rats were cultured for 24 hr with WsEV, SsEV, or vehicle. There was no difference in particle distribution and total concentration between WsEV and SsEV. Both SsEV and WsEV had no significant effect on the KCl-induced maximal contraction, while SsEV specifically attenuated contraction induced by noradrenaline compared with WsEV- and vehicle-treatment. In summary, it was for the first time revealed that SsEV attenuate the agonist-induced contractility of isolated blood vessels, which might be at least partly responsible for the BP regulation by SsEV.

Polyunsaturated fatty acids including arachidonic acid (AA), docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA), are converted to lipid mediators by oxidation. Unlike other mammals, cats cannot synthesize AA. Since their lipid metabolic features remain unknown, we qualitatively analyzed 118 types of urinary lipid metabolites in healthy neutered cats. Using LC-MS, we found 26 lipid metabolites in urines of all individuals. In detail, 20 AA-, 5 EPA- and 1 DHA-derived lipid mediators were detected. Focusing on oxidative pathway, 17 cyclooxygenase-metabolites and 5 metabolites produced by non-enzymatic pathway were detected. Of interest, few lipoxygenase- or cytochrome P450-metabolites were excreted. Thus, AA-derived cyclooxygenase-metabolites mainly composed the urinary lipid metabolites in cats.

To evaluate the effect of antimicrobial susceptibility on outcomes, we compared the minimum inhibitory concentrations (MICs) for Staphylococcus, Streptococcus, and the family Enterobacteriaceae from cured and uncured mastitis cases; milk shipment for uncured cases could not be resumed within 3 weeks after initial clinical examination. A higher MIC₅₀ of ampicillin and a higher MIC₉₀ of cefazolin for Enterobacteriaceae isolates were observed for cured rather than uncured cases with differences in ≥2 tubes. Endotoxins are generally released from Enterobacteriaceae upon antimicrobial treatment; their amounts are presumed to be greater in mastitis cases resulting from β-lactam antibiotic-susceptible rather than -resistant microbes. For staphylococcal and streptococcal isolates, the MIC₅₀ and MIC₉₀ of β-lactam antibiotics were similar for cured and uncured cases.

The object of this study is to evaluate the long-term outcome of hind limb weight-bearing function and progression of stifle osteoarthritis (OA) after tibial plateau leveling osteotomy (TPLO). Groups were classified by the degree of cranial cruciate ligament (CrCL) damage and presence or absence of medial meniscus damage as macroscopically evaluated during surgery. Weight-bearing function was assessed via the peak vertical force (PVF), and OA progression was assessed via the radiographic OA score (OAS) preoperatively and 1, 3, 6, 12, 18, 24, and 36 months postoperatively. In all stifles, PVF was significantly higher within 6 months postoperatively than preoperatively, and this high ratio was maintained for 36 months. The OAS was significantly higher 24 months postoperatively than preoperatively in stifles with a partial CrCL tear, and significantly increased at each timepoint after 3 months postoperatively in stifles with complete CrCL rupture. The OAS remained consistently lower in stifles with a partial CrCL tear and no treatment of the medial meniscus than in stifles with complete rupture. Thus, after TPLO, the weight-bearing function improved in the early postoperative period and was maintained for as long as 36 months. Although OA progressed over time after TPLO, the progression was more gradual in stifles with partial tears than in those with complete rupture. Canine patients would benefit from earlier surgical intervention through development of technology that enables early detection of ligament degeneration.

To evaluate the sedative and physiological effects of alfaxalone intramuscular (IM) administration, 12 healthy cynomolgus monkeys were administered single IM doses of alfaxalone at 0.625 mg/kg (ALFX0.625), 1.25 mg/kg (ALFX1.25), 2.5 mg/kg (ALFX2.5), 5 mg/kg (ALFX5), 7.5 mg/kg (ALFX7.5), or 10 mg/kg (ALFX10); saline was used as the control (CONT). The sedative effects were subjectively evaluated using a composite measure scoring system in six animals. Changes in respiratory rate, pulse rate, non-invasive blood pressure, percutaneous oxygen-hemoglobin saturation (SpO₂), and rectal temperature were observed after IM treatments in the other six animals. All animals were allowed to lay down following the ALFX5, ALFX7.5, and ALFX10 treatments, whereas lateral recumbency was achieved in only two animals after ALFX2.5 treatment and none after the CONT, ALFX 0.625, and ALFX1.25 treatments. The median time (interquartile range) to lateral recumbency was 6.5 min (5.3–7.8), 4.0 min (4.0–4.0), and 3.0 min (3.0–3.8), and the duration of immobilization was 27.5 min (19.0–33.8), 56.0 min (42.3–60.8), and 74.5 min (62.8–78.0) after the ALFX5, ALFX7.5, and ALFX10 treatments, respectively. Endotracheal intubation was achieved in all six animals after the ALFX7.5 and ALFX10 treatments. Dose-dependent decreases in respiratory rate, non-invasive blood pressure, SpO₂, and rectal temperature were observed, and the quality of recovery was smooth in all animals after the ALFX5, ALFX7.5, and ALFX10 treatments. Thus, alfaxalone IM induced a dose-dependent sedative effect in cynomolgus monkeys, but at higher doses, hypotension, hypoxemia, and hypothermia could be induced.

A 1-year-old male mixed breed dog presented for the evaluation of progressive hindlimb paresis. Neurological examination indicated a spinal cord lesion between the 3rd thoracic and 3rd lumbar vertebrae. Magnetic resonance imaging (MRI) revealed an intramedullary spinal cord lesion located at the level of the 1st and 2nd lumbar vertebrae. Following cytoreductive surgery of the mass, palliative radiation therapy was administered. A diagnosis of nephroblastoma was made based on histological examination. After radiation therapy, the disappearance of the spinal lesion was confirmed by MRI. The dog was improved from gait abnormality and alive at 16 months postoperatively, with slight signs of neurological dysfunction.

The purpose of this study was to investigate the effects of pupil diameter on canine visual evoked potentials with pattern stimulation (P-VEP). Atropine eye drop (1.0%) was applied to both eyes as a cycloplegic drug, and tafluprost eye drop (0.015%) was applied to one eye that was selected randomly for miosis (miosis group). The other eye did not receive tafluprost (mydriasis group). P-VEP was recorded at three pattern sizes. The P100 implicit time at a small pattern size in the mydriasis group was significantly prolonged compared to the miosis group. We hypothesized that the prolonged P100 implicit time under mydriatic conditions was due to increased spherical aberrations and concluded that mydriatic conditions affected P100 implicit time in canine P-VEP recordings.

A lamb presented with recurrent prolapse of the descending colon. On clinical examination, intussusception of the descending colon with the prolapse of a segment was verified. The external anal sphincter had a rupture, extending to the lacerated wound in the anus. The lamb underwent colopexy with the two-portal video-assisted incisional technique and was discharged 6 days after the surgical procedure with a satisfactory clinical outcome. There were no recurrences or complications for at least 9 months. Video-assisted colopexy is an alternative treatment for intussusception and recurrent colon prolapse in sheep, even in the presence of an external anal sphincter rupture.

This study aimed to describe the duration of inflammation after intrauterine infusion of polyvinylpyrrolidone-iodine (povidone-iodine, PVP-I), determine the effect of PVP-I infusion on the subsequent fertility, and evaluate the histopathology of the endometrium in dairy cows. In Experiment 1, 120 lactating clinically healthy Holstein-Friesian cows at 5 weeks postpartum (W5) were equally divided into three groups: intrauterine infusion of 2% PVP-I (PVP), saline (SAL), and no treatment (NTX). Endometrial cytology was performed daily from D0 (W5) to D7 to determine the percentage of polymorphonuclear cells (PMN%) in 44 of the 120 cows. All cows received timed artificial insemination at D17. In Experiment 2, 25 cows were randomly classified into sacrifice at 24 hr or 48 hr after 2% PVP-I infusion (PVP24 and PVP48), and 24, 48, 72, or 96 hr after SAL infusion (SAL24; SAL48; SAL72; SAL96), or no treatment (NTX). Histopathology was performed on the uterus of each cow. In Experiment 1, PMN% was greater in PVP (P<0.05) than in SAL and NTX, on D1, but decreased to a level similar to that of the other groups by D2. Conception rate was higher (P<0.05) in PVP cows compared to SAL and NTX cows. In Experiment 2, stratified columnar epithelium in the uterus disappeared in PVP24 and SAL24. The epithelium was regenerated in PVP48, SAL72, and SAL96, but not in SAL48. In conclusion, the results of the study suggest that PVP-I induces transient uterine inflammation, promotes regeneration of endometrial epithelial cells and improves fertility.

A high-concentrate diet destroys gram-negative bacteria in the cattle rumen, leading to elevated ruminal lipopolysaccharide (LPS) levels. LPS causes liver inflammation through the hepatic portal vein but little is known about the effects of rumen-derived LPS on liver function and the reproductive organs. In this study, we determined the effect of increasing rumen fluid LPS levels on liver function and genital LPS levels. Cows were assigned to control (CON; n=5) and high-concentrate diet (HC; n=7) groups. We observed that the ruminal LPS and haptoglobin (Hp) levels were significantly higher and albumin levels were lower in the HC group than in the CON group. In the HC group, The Hp levels and aspartate transaminase (AST) activity were significantly higher and the total cholesterol levels were significantly lower after high-concentrate diet feeding than before feeding. No differences were observed in LPS levels in the peripheral veins, hepatic veins, hepatic portal vein, uterine perfusate, and follicular fluids between the groups. In all samples, the LPS level in the hepatic portal vein blood positively correlated with the AST activity and serum amyloid A level. In conclusion, our results indicate that high-concentrate diets do not have a direct effect on the reproductive organs upon a moderate ruminal LPS level increase. However, an increased ruminal LPS influx into the liver might affect negatively liver function.

The knowledge of cytochrome P450 (CYP) expression involved in chemical exposure are necessary in clinical applications for the medication and prediction of adverse effects. The aim of this study was to evaluate the mRNA expression of CYP1–CYP3 families in cats exposed to BDE-209 for one year. All selected CYP isoforms showed no significant difference in mRNA expressions between control and exposure groups, however, CYP3A12 and CYP3A131 revealed tend to be two times higher in the exposure group compared to control group. The present results indicate that the chronic exposure of BDE209 could not alter CYP expression in the liver of cats. This result considered caused by the deficiency of CYP2B subfamily which is major metabolism enzyme of polybrominated diphenyl ethers (PBDEs) in cat.

The objective of the present study was to evaluate the cross-protective immunity between type 1 and type 2 porcine reproductive and respiratory syndrome virus (PRRSV) isolates in growing pigs. Japanese type 1 PRRSV, first isolated from a pig with respiratory disorders in a farm in 2009, exhibits unique genetic characteristics. The pathogenicity of a Japanese standard strain of type 2 PRRSV, EDRD1, in pigs immunized by the type 1 PRRSV isolate, Jpn EU 4-37 was determined by evaluating clinical signs, viremia, antibody response, and pathological lesions. Similarly, we evaluated the pathogenicity of Jpn EU 4-37 in pigs immunized by EDRD1 and compared the cross-protective immunity between these isolates. The EDRD1 challenge after Jpn EU 4-37 inoculation reduced viral clearance and shedding in pigs, compared to those treated with the EDRD1 single infection. On the other hand, the pathogenicity of Jpn EU 4-37 after EDRD1 infection did not differ significantly compared to non-immunized pigs treated with Jpn EU 4-37. Therefore, exposure to Jpn EU 4-37 could not induce enough immunity to reduce the viremia against subsequent infection by type 2 PRRSV. However, the immunity induced by Jpn EU 4-37 infection may play a role in reducing viremia caused by type 2 PRRSV. Moreover, the immunity induced by the EDRD1 and other genetically related viruses, which are broadly distributed in Japan, may not contribute to cross-protection against Jpn EU 4-37 as an emerging virus.

Mosquitoes transmit many kinds of arboviruses (arthropod-borne viruses), and numerous arboviral diseases have become serious problems in Indonesia. In this study, we conducted surveillance of mosquito-borne viruses at several sites in Indonesia during 2016–2018 for risk assessment of arbovirus infection and analysis of virus biodiversity in mosquito populations. We collected 10,015 mosquitoes comprising at least 11 species from 4 genera. Major collected mosquito species were Culex quinquefasciatus, Aedes albopictus, Culex tritaeniorhynchus, Aedes aegypti, and Armigeres subalbatus. The collected mosquitoes were divided into 285 pools and used for virus isolation using two mammalian cell lines, Vero and BHK-21, and one mosquito cell line, C6/36. Seventy-two pools showed clear cytopathic effects only in C6/36 cells. Using RT-PCR and next-generation sequencing approaches, these isolates were identified as insect flaviviruses (family Flaviviridae, genus Flavivirus), Banna virus (family Reoviridae, genus Seadornavirus), new permutotetravirus (designed as Bogor virus) (family Permutotetraviridae, genus Alphapermutotetravirus), and alphamesoniviruses 2 and 3 (family Mesoniviridae, genus Alphamesonivirus). We believed that this large surveillance of mosquitoes and mosquito-borne viruses provides basic information for the prevention and control of emerging and re-emerging arboviral diseases.

Bovine leukemia virus (BLV) belongs to the genus, Deltaretrovirus of the family, Retroviridae and it is the causative agent of enzootic bovine leukosis. The prevalence of BLV in three provinces in the Red River Delta Region in the North of Vietnam, Hanoi, Vinhphuc and Bacninh was studied from April 2017 to June 2018. A total of 275 blood samples collected from cattle were used for serum isolation and DNA extraction. Of these samples, 266 sera were subjected to ELISA test for detecting antibody against BLV gp51 protein and 152 DNA samples were used to detect the 444 bp fragment corresponding to a part of the gp51 region of the env by nested PCR. The results showed that 16.5% (n=44) and 21.1% (n=32) of samples were positive for BLV gp51 antibody and BLV proviral DNA, respectively. Phylogenetic analysis of the partial (423 bp) and complete (913 bp) BLV env-gp51 gene indicated that Vietnamese strains were clustered into genotypes 1, 6 and 10 (G1, G6 and G10). Of those genotypes, G1 genotype was dominant; G6 strains were designated as G6e and G6f subgenotypes; the existence of genotype 10 was confirmed for the first time in Vietnam. The present study provides important information regarding the prevalence of BLV infection and genetic characteristics of BLV strains identified in Vietnam, contributing to promote the establishment of disease control and eradication strategies in Vietnam.

The anthropometric and blood data of an unsuccessfully hand-reared Asian elephant (Elephas maximus) calf were retrospectively compared with the data for calves raised by their real mothers or allomothers, to identify potential reasons for poor outcomes in the hand-reared case. The hand-reared calf grew normally in terms of body weight and withers height. However, blood biochemical data suggested reduced bone metabolism, low immune status, and malnutrition during its life. Blood bone markers were measured to determine whether a skeletal disorder was present in the Asian elephant calf, which was not clear from the anthropometric data. Monitoring these parameters in hand-reared Asian elephant calves, with the aim of keeping them within the normal range, may increase the success rate of hand-rearing of Asian elephant calves.

The prevention of diseases through health control is essential at zoos. Here, we investigated the gut microbiota formation during infancy in an Asian elephant and compared the composition between infant and mother. Besides, we analyzed the components of breast milk and examined the correlation with the infant gut microbiota. Analysis revealed the gut microbiota of the infant contained high amount of Lactobacillales and its diversity was relatively low compared to that of the mother. We found several milk components, showed a positive correlation with the change of Lactobacillales. The present study revealed the mechanism of gut microbiota formation during infancy in an Asian elephant and provides important insights into the health control of Asian elephants in zoos.