Lysophosphatidic acid (LPA) produced by autotaxin (ATX) is recognized as a multi-functional mediator in mammalian reproduction. This study focused on possible effect(s) of LPA on ovulated cumulus-oocyte complexes (COCs) around fertilization in rats in vivo. Immunohistochemistry revealed the cell-type-dependent localization of candidates of synthetic enzymes, ATX and two phospholipases A2 isofroms, and LPA receptors LPA1~4 in ovulated COCs and in oviductal epithelium. The eggs ovulated with a form of COCs became denuded of cumulus cells and underwent fragmentation in the absence of fertilization. In vivo experiments of local administration in non-copulated rats demonstrated that eggs denudation was increased by LPA and decreased by anti-ATX antibody and that fragmentation was inhibited by LPA and stimulated by an ATX chemical inhibitor. Furthermore, LPA administration in adult copulated rats increased the rate of cleaved embryos significantly. Obtained results suggest the presence of LPA synthesis and action system in ovulated COCs within the oviductal ampulla and positive actions of LPA possibly at multiple sites around fertilization in rats.
Gliomas are common intracranial neoplasias in dogs. However, the underlying pathogenic mechanisms remain unclear. In humans, isocitrate dehydrogenase 2 (IDH2) is often mutated in gliomas. Although almost human IDH2 mutations have been identified at the Arg172 codon, few studies have reported structural, functional, or mutational information for canine IDH2. In this study, we cloned the full-length canine IDH2 (cIDH2) cDNA and substituted wild type Arg174 (cIDH2 WT: corresponding to R172 of human IDH2) with Lys (cIDH2 R174K). The cIDH2 WT and R174K proteins were overexpressed in HeLa cells, and their presence was confirmed using an anti-human IDH2-WT mAb (clone: KrMab-3) and an anti-IDH2-R172K mAb (clone: KMab-1). The IDH2 activity between cIDH2 WT and cIDH2 R174K transfectants was compared by measuring the production of NADH and NADPH. NADPH production was lower for cIDH2 R174K than that for cIDH2 WT transfectants. Finally, we detected increased expression of hypoxia inducible factor-1 alpha (HIF-1α) in cIDH2 R174K transfectants. This indicates that mutations at R174 can potentially induce carcinogenesis in canine somatic cells.
A cross-sectional study was conducted to estimate the prevalence of Fasciola hepatica (F. hepatica) infection in water buffalo (Bubalus bubalis) in Alexandria, Beheira, and Kafr el-Sheikh governorates (provinces) of the Nile Delta in Egypt and to identify the underlying risk factors associated with the infection. A total of 29 farms (10 in Alexandria, 10 in Beheira, and 9 in Kafr el-Sheikh) were randomly selected and all the buffaloes that resided on these farms from 21 February 2015 to 20 February 2016 were included in the study. The sampling approach was target-based where all the buffaloes were examined and screened for clinical signs of Fasciola infection. All suspected buffaloes were then subjected to fecal examination, and those positive for Fasciola eggs underwent antibody testing using indirect hemagglutination test. Consequently, data on 3,356 buffaloes from 29 farms in these governorates was analyzed using a multiple logistic regression model. The final model showed that the age and body condition score of the buffalo, location and type of the farm, application of prophylactic treatment, and temperature and relative humidity of the farm’s location significantly affected the rate of infection. The highest prevalence was observed in buffaloes from Alexandria governorate (19.6%), followed by Beheira and Kafr el-Sheikh governorates (15.5% and 9.1%, respectively).
Campylobacterjejuni and C. coli are the leading causes of enteric infections in many developed countries. Healthy chickens are considered to act as reservoirs of campylobacters, as the organisms colonize the intestinal tract. Once infected birds enter a processing plant, contamination of chicken carcasses with campylobacters occurs over the entire skin during defeathering and evisceration due to leakage of crop and/or intestinal contents. Although the role of feather follicles in the contamination of chicken carcasses by campylobacters during processing is still debatable, it has been considered that the microorganisms would be entrapped and retained in the follicles due to the morphological changes resulting from defeathering and chilling. In the present study, we observed the morphology of feather follicles in chicken carcasses after defeathering and chilling. A total of 3,133 feather follicles were examined for morphological changes before and after chilling. Shortly after defeathering, most (91.5%) of the follicles were closed, whereas after chilling they were either closed (85.5%) or open (6%), although a small proportion of enlarged follicles became smaller or closed (2.6%). Moreover, 5.9% of the follicles that were slightly open became further enlarged after chilling. Furthermore, the proportion of enlarged feather follicles that became closed after chilling showed no discernible relationship with the degree of campylobacter contamination in different areas of the carcass skin, suggesting that campylobacters may not be confined to feather follicles as a result of the morphological changes attributable to defeathering and chilling.
The blood biochemical analysis of bone markers could have a role in the early diagnosis of metabolic bone disease in animals; however, there is limited information on bone markers in captive Asian elephants (Elephas maximus). Serum samples from ten captive Asian elephants were obtained to clarify the relationship between age and the blood bone markers tartrate-resistant acid phosphatase isoform 5b (TRAP5b) and bone specific alkaline phosphatase (BALP). Serum TRAP5b and BALP activities were negatively correlated with age. A positive correlation was observed between TRAP5b activity and BALP activity. These results may contribute to the health management of captive Asian elephants.
Atlanto-axial (AA) instability due to ligament insufficiency is a common cause of cervical spinal cord compression in toy breeds. However, in some dogs a difference in size between the atlas and the axis leads to joint incongruence that exacerbates AA subluxation and makes surgical treatment challenging. Twelve dogs with AA instability with incongruence were enrolled in a single institution prospective observational study. Computed tomography (CT) and magnetic resonance imaging (MRI) of the AA joint were compared to a retrospectively reviewed control group. A novel surgical approach consisting of a dorsal internal fixation technique was performed in six dogs. For affected dogs, the mean normalised difference between the dorso-ventral atlas canal and the dorso-ventral axis canal was 29.67% (median of 35.07%, standard deviation 25.64%), while in normal dogs a mean difference of 4.67% (median of 3.95%, standard deviation 5.21%) was observed. On MRI, 12/12 affected dogs had spinal cord compression, which was classified as reducible (3/12), partially reducible (6/12) and non-reducible (3/12). In surgically operated dogs, follow-up CT showed a partial or complete reduction of the previous spinal cord compression with a consistent amelioration or resolution of the presenting complaints. The proposed surgical technique was safe and effective in dogs with partially or completely reducible spinal cord compression.
In the swine industry, Lawsonia intracellularis is one of the main enteric pathogens; it causes acute intestinal hemorrhage (proliferative hemorrhagic enteropathy) in naïve adult pigs and a wasting disease (proliferative enteropathy) in growing pigs. Among many kinds of cytokines, interferon-γ (IFN-γ) has previously been reported to play a significant role in limiting intracellular infection and increasing cellular proliferation associated with L. intracellularis. However, the levels of various circulating inflammatory cytokines, including IFN-γ, in animals infected with L. intracellularisis is still an area of considerable interest for understanding immunity against this bacterium. In addition, there has been no information on cytokine response in animals infected with any L. intracellularis isolate of South Korean origin or Asian origin. To determine the relationship between the changes in the systemic inflammatory cytokine response in the peripheral blood of the host after L. intracellularis infection, we measured the levels of some pro-inflammatory cytokines such as tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and IFN-γ, anti-inflammatory cytokines (IL-4, IL-10, and transforming growth factor-β (TGF-β)), and a chemokine (IL-8) in pigs infected with L. intracellularis isolated from South Korea. This study demonstrated that a L. intracellularis isolate of South Korean origin induced cytokine (TNF-α, IL-6, and IFN-γ) responses in infected animals within 15 days post-infection although the circulating levels of IL-4, IL-10, IL-8 and TGF-β were induced relatively late.
Vibrio vulnificus is known as an opportunistic bacterial pathogen that causes primary septicemia and wound infection in humans. Recently, the incidence of wound infection by V. vulnificus is increasing in warm countries. In this study, we examined a vaccine antigen against V. vulnificus in mice. FlaB, a component protein of the V. vulnificus flagellum, was expressed as a recombinant protein, named rFlaB. After immunization of mice with rFlaB, the mice were challenged by subcutaneous inoculation with V. vulnificus. Bacterial burdens in muscular tissue at the infection site in rFlaB-immunized mice were significantly decreased compared with those of control mice. We found that rFlaB immunization can partially suppress proliferation of V. vulnificus at the local infection site.
Two dogs presented to the emergency service after accidental ingestion of afloqualone tablets, a muscle relaxant used for back pain in humans. Toxic effects of the drug in these dogs included vomiting, respiratory depression, seizures, ataxia, bradycardia, and hematuria. Treatment consisted of fluid diuresis, furosemide, and propofol. Flumazenil, a gamma-amino butyric acid antagonist, was administered intravenously; however, it was not effective in stopping the seizures in these dogs. Both dogs recovered with supportive treatment. To the authors’ knowledge, this is the first documented report of afloqualone intoxication in dogs.
This study aims to evaluate the oxidative stress during hot summer season using serum oxidative stress biomarkers and elucidate the effects of serum antioxidant vitamin levels in dairy and beef cows in a daytime grazing system. Blood samples were collected once a month from eight Holstein Friesian (HF) and 10 Japanese Black (JB) cows from November 2013 to October 2014. Serum values of derivatives of reactive oxygen metabolites (d-ROMs) tended to be higher in March in both breeds and those in HF cows were kept at higher (P<0.001) levels than those in JB cows during the study period. Serum levels of biological antioxidant potential (BAP) in both breeds were maintained at almost the same values during study period. The OSI [(d-ROMs/BAP) × 100] values in both breeds showed similar seasonal changes, i. e. increase from December to March and decrease from March to August or September. In addition, the OSI values in HF cows were kept at higher (P<0.01) levels than those in JB cows during the study period. Serum concentrations of α-tocopherol, β-carotene, blood urea nitrogen and total cholesterol showed similar seasonal changes in both breeds, low in the winter and high from spring to summer, which may be attributed to the pasture grass intake. Opposite changes in OSI values and serum concentrations of α-tocopherol and β-carotene indicated that antioxidant vitamin levels could affect oxidative stress status.
Liposarcoma of the spermatic cord is extremely rare in dogs and humans. This report describes the clinical signs, typical diagnostic imaging including ultrasound and computed tomography, and treatment of a liposarcoma of the spermatic cord of a Toy Poodle confirmed by histological examination after a surgical procedure. This case highlights the importance of preoperative diagnostic imaging and histopathological examination in dogs with an inguinal or scrotal mass.
This study was performed to evaluate the effect of maternal supplementation with seaweed powder (SWP) on the immune status of piglets. Sows were supplementary fed SWP from 85-days of gestation until delactation. Forty-days old piglets were euthanized and lymphocyte subsets were analyzed. The results showed a significantly higher relative population of CD4+CD8+ T cells in the thymus, lymph node, tonsil (P<0.05), peripheral blood mononuclear cells, spleen and liver (P<0.01) of piglets derived from treated sows. A higher relative population of CD8+ T cells was also observed in the liver and spleen (P<0.05) of the piglets. The data suggested the enhancing effects of maternal supplementation with SWP on immune status of piglets.
Dynamics of serum liver enzymes in rabbits experimentally infected with metacercariae of Fasciola sp. (intermediate form between Fasciola hepatica and F. gigantica) were monitored. Gradual increase of serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were observed from 3 weeks post-inoculation (WPI) and peaked at 6 WPI, which corresponded well to the period of migration and development of juvenile fluke in the liver parenchyma and the time when the young adult flukes migrated to the bile duct. However, no significant increase in serum gamma-glutamyl transferase (GGT) and alkaline phosphatase (ALP) were observed. This could reflect reduced or minimal injury of bile ducts and biliary epithelia as the flukes had reached the adult stage. Alpha- fetoprotein (AFP) and carcinoembryonic antigen (CEA) were not detected in the infected rabbit during the course of the experiment. Serum liver enzymes monitoring might be useful for understanding the host-parasite relationship in fascioliasis.
To investigate the utility of cerebrospinal fluid (CSF) anti-feline coronavirus (FCoV) antibody test for diagnosis of feline infectious peritonitis (FIP), the antibody titers were tested in CSF and sera from 271 FIP-suspected neurological cats. CSF antibody was detected in 28 cats, which were divided into 2 groups; 15 with CSF titer of 1:80 or lower and 13 with CSF titer of 1:640 or higher. In the latter group, reciprocal serum titer/reciprocal CSF titer was 8 or lower, which is extremely lower than normal range (256-2048), and FCoV RNA was detected in all of 11 CSF samples assayed by RT-PCR. Our findings indicate that CSF titer of 1:640 or higher may be served as a candidate for the index for diagnosing FIP.
Haemorrhagic septicemia (HS) is a contagious disease in cattle with high morbidity and mortality rates. HS vaccine in Thailand is an oil-adjuvant formulation, and is difficult to administer. The present study aimed to formulate and evaluate the protection in dairy calves conferred by immunization with an in-house intranasal HS vaccine. The intranasal vaccine was formulated in a total volume of 500 µl containing either 50 or 100 µg of the recombinant outer membrane protein H (rOmpH) of Pasteurella multocida strain M-1404 (serovar B:2), and 10 µg of Cytosine-phosphate-guanosine oligodeoxynucleotides (CpG-ODN) as a mucosal adjuvant. Intranasal immunizations were conducted three times at three-week intervals. The antibodies post-immunization were detected by indirect ELISA and demonstrated efficient in vitro activity in suppressing a P. multocida strain from the complement-mediated killing assay. An intranasal vaccine induced both the serum IgG and secretory IgA levels that were significantly higher than the level conferred by the parenteral vaccine (P<0.05). Challenge exposure was conducted with a P. multocida strain M-1404 at day 72 of the experiments. The immunized calves had reduced clinical signs after challenge exposure that would normally result in disease proliferation. We conclude that intranasal vaccination of calves with rOmpH with CpG-ODN 2007 stimulated serum and secretory antibodies to rOmpH and whole cells of P. multocida strain M-1404 antigen. Moreover, it would result in protection in calves against artificial P. multocida infection.
The embryonic diaphragm comprises four major structural components derived from the transverse septum, the dorsal foregut mesentery, the pleuroperitoneal folds (PPFs), and the body wall. In this study, the appearance of PPFs and related factors were investigated using light microscopy of horizontal sections of rat fetuses from embryonic day 12 to 13. In rat fetuses, the sign of PPF projection was noted in the sidewall of the pericardioperitoneal canal at embryonic day 12, and was confirmed as folds at embryonic day 12.25. Expressions of GATA4, COUP-TF2, and FOG2 were detected in PPF at the early stage of formation. Localizations of these factors suggested that COUP-TF2 and FOG2 are the main factors in PPF appearance and that GATA4 is unlikely to be a main factor, although it is necessary for PPF formation.
The sequence of cranial suture closure among cervids is reported to be generally species-specific and highly conservative within species. On the other hand, it is known that intraspecific variation often exists to some extent in other mammalian taxa. Here we studied the cranial suture closures of Capreolus pygargus from Jeju Island and compared it with other cervid species. We found that the timing of the interparietal suture closure is highly variable within C. pygargus. Capreolus capreolus similarly shows intraspecific variation of the interparietal suture closure, whereas other cervid species studied to date do not show any intraspecific variation in the sequence of cranial suture closure. Such high intraspecific variation of the interparietal suture may be a derived character for Capreolus.
Enzootic bovine leukemia is caused by the bovine leukemia virus (BLV). BLV is transmitted vertically or horizontally through the transfer of infected cells via direct contact, through milk, insect bites and contaminated iatrogenic procedures. However, we lacked direct evidence of intrauterine infection. The purpose of this study was to confirm intrauterine BLV infection in two pregnant dams with high viral load by cesarean delivery. BLV was detected in cord and placental blood, and the BLV in the newborns showed 100% nucleotide identity with the BLV-env sequence from the dams. Notably, a newborn was seropositive for BLV but had no colostral antibodies. In this study, we presented a direct evidence of intrauterine BLV transmission in pregnant dam with a high proviral load. These results could aid the development of BLV control measures targeting viral load.
The aim of this study was to investigate osteoclastogenic properties of inflammatory cytokines at different time-points of osteoclastogenesis. Bone marrow-derived macrophages from five healthy dogs were stimulated with the macrophage colony-stimulating factor, receptor activator of nuclear factor-κB ligand and inflammatory cytokines such as interleukin (IL)-1β, tumor necrosis factor (TNF)-α and IL-17. Osteoclasts (OC) formation and function were enhanced with TNF-α regardless of temporal differences. But in contrast, IL-1β suppressed the osteoclastogenesis at early phase of the process while upregulating at the late phase. Furthermore, differentiation of OC precursors into OC was suppressed at high concentrations of IL-17. Collectively, the results revealed that suppressing TNF-α would be a promising strategy to inhibit inflammation-associated bone destruction in dogs.
Bovine mastitis causes significant economic losses in the dairy industry. Effective prevention of bovine mastitis requires an understanding of the infection status of a pathogenic microorganism in a herd that has not yet shown clinical signs of mastitis and appropriate treatment specific for the pathogenic microorganism. However, bacterial identification by culture has drawbacks in that the sensitivity may be low and the procedure can be complex. In this study, we developed a genetic detection method to identify mastitis pathogens using a simple and highly sensitive electrochemical DNA chip which can specifically detect bacterial DNA in milk specimens. First, we selected microorganisms belonging to 12 families and/or genera associated with mastitis for which testing should be performed. Next, we optimized the conditions for amplifying microorganism DNA by loop-mediated isothermal amplification (LAMP) using 32 primers and the use of a DNA chip capable of measuring all pathogens simultaneously. Sample detection could be completed in just a few hours using this method. Comparison of the results obtained with our DNA chip method and those obtained by bacterial culture verified that when the culture method was set to 100%, the total positive concordance rate of the DNA chip was 85.0% and the total negative concordance rate was 86.9%. Furthermore, the proposed method allows both rapid and highly sensitive detection of mastitis pathogens. We believe that this method will contribute to the development of an effective mastitis control program.
Urethral prolapse is a rare condition in dogs and cats. A 7-month-old male cat presented with intermittent penile bleeding and a red mass protruding from the tip of the penis. Urethral prolapse was diagnosed by using a surgical microcamera to examine the genital area. Urethroplasty and preputial advancement surgery were performed using a surgical microscope, to successfully correct the urethral prolapse.
A prospective observational study was performed in canine clinical medicine to evaluate the emetic action and adverse effects of tranexamic acid. Veterinarians treated 137 dogs with a single dose of tranexamic acid (50 mg/kg, IV) after accidental ingestion of foreign substances. If needed, a second (median, 50 mg/kg; range, 20–50 mg/kg, IV) or third dose (median, 50 mg/kg; range, 25–50 mg/kg, IV) was administered. Tranexamic acid induced emesis in 116 of 137 (84.7%) dogs. Median time to onset of emesis was 116.5 sec (range, 26–370 sec), median duration of emesis was 151.5 sec (range, 30–780 sec), and median number of emesis episodes was 2 (range, 1–8). Second and third administrations of tranexamic acid induced emesis in 64.7% and 66.7% of dogs, respectively. In total, IV administration of tranexamic acid successfully induced emesis in 129 of 137 (94.2%) dogs. Adverse effects included a tonic-clonic convulsion and hemostatic disorder in two different dogs, both of which recovered after receiving medical care. Tranexamic acid induced emesis in most dogs following a single-dose. When a single dose was not sufficient, an additional dosage effectively induced emesis. Overall, adverse effects were considered low and self-limiting.
A 16-year-old female Indian peafowl (Pavo cristatus) died two days after recognition of conjunctivitis in the right eye, anorexia and depression. Gross necropsy revealed a thick pseudomembrane under the eyelid and hydropericardium. Histopathological examination revealed hepatocellular necrosis, sinusoidal and vascular congestion and infiltrated inflammatory cells. Infiltration by inflammatory cells was noted in the epicardium. The lungs had mild interstitial pneumonia with the extensive congestion within the capillaries of the air sacs. Tubular interstitial congestion and necrosis was noted in the kidneys. Bacterial culture and nucleotide sequencing of the inflammatory specimens identified the causative agent as Serratia marcescens, an uncommon bacterium in birds. In summary, this study describes the sudden death of an Indian peafowl due to S. marcescens infection, which is rarely seen in animals.
A 2- to 4-year-old uncastrated male Siberian tiger (Panthera tigris altica) bred in a local wild animal park presented with generalized clinical signs including abdominal pain, fever, lethargy, and anorexia, along with subcutaneous nodules along the trunk. The patient subsequently died of chronic, progressive dyspnea despite 45 days of antibiotic treatment. At necropsy, mesenteric fat inflammation and multiple subcutaneous, peritoneal, and intraabdominal nodules were observed. The lungs demonstrated congestion and heavy coagulation, and necrotic foci were observed on the cut surface. Histopathologically, the nodules were identified as granulomatous fatty tissue with numerous lymphocytes, infiltration with lipid-laden macrophages, and fibrosis. These changes were also noted in the lung. The etiology of this condition remains undetermined.
A Japanese Black calf, manifested by clinical signs of inability to stand and extending hind limbs after birth, was investigated clinically and pathologically. In the neurological inspection, gastrocnemius reflex and patellar tendon reflex of the right hind limb were hyperactive, and gastrocnemius reflex and crossed extension reflex of the left hind limb were also hyperactive. Magnetic resonance imaging (MRI) examination showed a cavity in the cord at the area of the third and fourth lumbar vertebrae in T2-weighted imaging. After the calf was euthanized, necropsy confirmed the swelling of the spinal cord at the third and fourth lumbar vertebrae. The histopathological examination confirmed that the central canal remained open, and the inner surface of central canal was lined with ependymal cells. Therefore, this calf was diagnosed to have hydromyelia of the lumbar cord. This study indicated that MRI was useful for diagnosing myelodysplasia such as hydoromylia. This is the first report of hydoromylia in Japanese Black calf.
The role of cardiac autonomic modulation on ventricular arrhythmia, known as ventricular premature complexes (VPC), after splenectomy was investigated. Twelve dogs undergoing splenectomy were divided into 2 groups: low VPC (<1,000/day, n=6) and high VPC groups (≥1,000/day, n=6). Electrocardiograph recording was performed prior to (D0), during the first three days (D1–3) and on day 9 (D9) after surgery. Arrhythmic indices, Tpeak-Tend , corrected QT interval and short-term variability of QT interval as well as heart rate variability (HRV) were evaluated. Plasma concentrations of norepinephrine (NE) and epinephrine (E) were measured. In the high VPC group, the occurrences of VPC were significantly increased (P<0.05) after surgery, and reached the levels higher than those in the low VPC group. For the arrhythmic indices, only Tp-Te in the high VPC group increased significantly (P<0.05) after surgery. For HRV analysis, enhancement of both time and frequency domains were found postoperatively in both groups. On D2, however, the high VPC group showed significantly lower total power and high frequency with higher low to high frequency ratio (P<0.05) than the low VPC group. Plasma NE concentration significantly increased in the high VPC group after surgery. Dogs in the high VPC group had shorter survival time than those in the low VPC group. In conclusion, dogs with imbalance cardiac autonomic modulation accompanied with high circulating NE concentration after splenectomy are prone to ventricular arrhythmia, which leads to short survival time.
The purpose of this study was to compare the concentration of trace elements in the plasma of sea turtles that inhabited the suburban (Okinawa Main Island, n=8) and the rural coast (Yaeyama Island, n=57) in Okinawa, Japan. Particle induced X-ray emission allowed detection of 20 trace and major elements. The wild sea turtles in the suburban coast in Okinawa were found to have high concentrations of Pb, Si, and Ti in the plasma when compared to the rural area but there were no significant changes in the Al, As, and Hg concentrations. These results may help to suggest the status of some elements in a marine environment. Further, monitoring the plasma trace and major element status in sea turtles can be used as a bio-monitoring approach by which specific types of elements found here could indicate effects that are related to human activities.
The fine structures of different tendons in various animals at different ages have been studied extensively to reveal their arrangement and growth patterns. However, knowledge of the microstructures of the growing tenocytes in the tendons of piglets is still lacking. Thus, we performed the first morphometric analysis to describe the characteristics of tenocytes in the metacarpal superficial digital flexor tendon of 0-, 10-, and 20-day-old piglets. In the present study, hydrochloric acid/collagenase digestion was applied to remove the interstitial connective tissue and obtain clear visualization of intact tenocytes and their cytoplasmic processes (Cp). Then, the morphometry of the tenocytes was investigated by optical and electron microscopy. The mean ± SE values of the fascicle area, number of tenocytes/fascicle, cell density, number of Cp/tenocyte, length of Cp, and thickness of Cp were compared among the three age groups. Significant differences (judged at P<0.05) were found in almost all morphometric aspects among the age groups, except for the number of Cp/cell (P=0.545) and thickness of the Cp (P=0.105). A decrease of cell density corresponded with an increase in the length of the Cp, which were extended to connect either with the Cp of the other tenocytes or the surrounding endotendineum. Moreover, an increase of the fascicle area reflected the increase in tendon diameter. The revealed morphometric characteristics are thus the outcome of tendon growth.
An indirect enzyme-linked immunosorbent assay (ELISA) using lipopolysaccharide extract as antigen was evaluated for detection of antibodies to Actinobacillus pleuropneumoniae serovar 15. The serovar 15 ELISA had a higher sensitivity and specificity than latex agglutination test for 63 and 80 sera from pigs experimentally infected and not infected with A. pleuropneumoniae, respectively. When the serovar 15 ELISA was applied to 454 field sera, high rates of seropositivity were found in pigs from farms infected with A. pleuropneumoniae serovar 15, but not in those from farms free of A. pleuropneumoniae serovar 15. The results suggest that the serovar 15 ELISA may be useful for the serological surveillance of infection with A. pleuropneumoniae serovar 15.
Anesthetic and cardiorespiratory effects of total intravenous anesthesia (TIVA) technique using propofol-guaifenesin-medetomidine (PGM) and alfaxalone-guaifenesin-medetomidine (AGM) were preliminarily evaluated in Thoroughbred horses undergoing castration. Twelve male Thoroughbred horses were assigned randomly into two groups. After premedication with intravenous (IV) administrations of medetomidine (5.0 μg/kg) and butorphanol (0.02 mg/kg), anesthesia was induced with guaifenesin (10 mg/kg IV), followed by either propofol (2.0 mg/kg IV) (group PGM: n=6) or alfaxalone (1.0 mg/kg IV) (group AGM: n=6). Surgical anesthesia was maintained for 60 min at a constant infusion of either propofol (3.0 mg/kg/hr) (group PGM) or alfaxalone (1.5 mg/kg/hr) (group AGM), in combination with guaifenesin (80 mg/kg/hr) and medetomidine (3.0 μg/kg/hr). Responses to surgical stimuli, cardiorespiratory values, and induction and recovery characteristics were recorded throughout anesthesia. During anesthesia induction, one horse paddled in group PGM. All horses from group AGM were maintained at adequate anesthetic depth for castration. In group PGM, 3 horses showed increased cremaster muscle tension and one showed slight movement requiring additional IV propofol to maintain surgical anesthesia. No horse exhibited apnea, although arterial oxygen tension decreased in group AGM to less than 60 mmHg. Recovery quality was good to excellent in both groups. In conclusion, TIVA using PGM and AGM infusion was available for 60 min anesthesia in Thoroughbred horses. TIVA techniques using PGM and AGM infusion provided clinically acceptable general anesthesia with mild cardiorespiratory depression. However, inspired air should be supplemented with oxygen to prevent hypoxemia during anesthesia.
For field-identification of taeniid cestodes in canine animals in Tibetan area, loop-mediated isothermal amplification (LAMP) assays for Echinococcus multilocularis, E. shiquicus, Taenia hydatigena, T. multiceps, T. pisiformis and T. crassiceps were developed and evaluated along with the reported assay for E. granulosus. The LAMP assays showed specific reaction with their corresponding target species DNA with the detection limit of 1 to 10 pg. Moreover, the assays for E. granulosus, E. multilocularis, T. hydatigena and T. multiceps could detect DNA extracted from 3 or more eggs of their corresponding target species. Then, the LAMP assays were applied on samples containing 3 to 35 taeniid eggs obtained from 61 field-collected canine feces in Qinghai, and the result was compared with a reported multiplex PCR and sequence analysis. The LAMP assays and the PCR detected single species DNA of E. granulosus, E. shiquicus, T. hydatigena and T. multiceps in 5, 2, 44 and 2 samples, respectively. In the rest 8 samples, DNA of both E. granulosus and T. hydatigena were detected by the PCR but the LAMP assays detected those DNAs in 2 samples and only T. hydatigena DNA in 6 samples. It was assumed that less than 3 E. granulosus eggs were mixed in the samples although the samples contained 21 to 27 eggs in total. In conclusion, the LAMP assays were less sensitive than the multiplex PCR, but would have adequate sensitivity for field use in Tibetan area.
The clinical and pathological records of 44 domestic, female rabbits with an age ranging from 6–124 months (median age: 63.5 month) were assessed retrospectively for ovarian lesions. Included were all rabbits that underwent an ovariohysterectomy with a subsequent pathological examination of the genital tract between March 1997 and June 2016. Pathological examination revealed ovarian lesions in 12 of the 44 rabbits including follicular cysts (n=7), cystic rete ovarii (n=3), widespread ovarian necrosis with dystrophic calcification (n=2), ovarian adenoma (n=1). Clinical examination including radiographs only suggested ovarian disorders in two cases of ovarian necrosis with dystrophic calcification and in two cases of cystic rete ovarii. Clinical significance was only conclusive in a case of cystic rete ovarii.
Sheep-associated malignant catarrhal fever (SA-MCF), caused by ovine gammaherpesvirus-2 (OvHV-2), is a fatal disease in all ruminants. The epidemiological survey and molecular characterization of OvHV-2 in Mongolian livestock were performed. Of 928 blood samples, 14 were positive for OvHV-2 in sheep and native cattle from Tsenkher County and in sheep from Lun County. Phylogenetic analyses revealed that the tegument gene of OvHV-2 sequences from Mongolian animals is identical to that in animals from Egypt, India, and Turkey, and is 98.0% similar to that in animals from Germany and Brazil. To our knowledge, this is the first confirmed report of OvHV-2 in Mongolian livestock, and could provide useful information for controlling SA-MCF.
Muscovy duck reovirus (MDRV) belongs to the Orthoreovirus genus of the Reoviridae family, which is a significant poultry pathogen leading to high morbidity and mortality in ducklings. However, the pathogenesis of the virus is not well understood. In the present study, two-dimensional (2D) polyacrylamide gel electrophoresis (PAGE) combined with LC-MS-MS was used to identify differentially expressed proteins between Muscovy duck embryo fibroblasts (MDEF) infected with virulent (MV9710 strain) and attenuated (CA strain) MDRV and non-infected MDEFs. A total of 115 abundant protein spots were identified. Of these, 59 of differentially expressed proteins were detected, with functions in metabolism and utilization of carbohydrates and nucleotides, anti-stress, and regulation of immune and cellular process. GO analysis of the identified proteins showed that they belonged to the classes molecular function (141 proteins), cellular component (62 proteins), and biological process (146 proteins). The results were validated by qRT-PCR, which suggests that the analysis method of 2D PAGE combined with LC-MS-MS used in this study is reliable. This study lays a foundation for further investigation of the biology of MDRV infection in MDEF.
Okayama University-type retinal prosthesis (OURePTM) is a photoelectric dye-coupled polyethylene film which generates electric potential in response to light and stimulates nearby neurons. This study aims to test surgical feasibility of subretinal implantation and functional durability of dye-coupled films in the subretinal space. The dye-coupled films were implanted subretinally by 25-gauge vitrectomy in the right eye of 11 normal beagle dogs: 2 dogs served for film removal after 5-month film implantation, 3 dogs for film removal after 3-month film implantation, 3 dogs for 3-month film implantation and pathological examination, and 3 dogs for sham surgery. The surface electric potential of the removed dye-coupled films in response to light was measured by the Kelvin Probe system. At surgery, rolled-up dye-coupled films in 5 × 5 mm square size could be inserted into subretinal space of retinal detachment induced by fluid injection with a 38-gauge polyimide tip. Retinal attachment was maintained by silicone oil injection in vitreous cavity. At autopsy, the retina in all dogs maintained the ganglion cell layer, inner and outer nuclear layers while it lost the outer segments in some part. All 5 sheets of removed dye-coupled films maintained the dye color. One sheet of the 5-month implanted film showed proportional increase of surface potential in response to increasing light intensity. Subretinal implantation of OURePTM by vitrectomy was technically feasible in canine eyes, and OURePTM maintained the function of generating light-evoked surface potential after 5 months in subretinal implantation.
Methylmercury (MeHg) is an environmental pollutant that shows severe toxicity to humans and animals. However, the molecular mechanisms mediating MeHg toxicity are not completely understood. We have previously reported that the MARCKS protein is involved in the MeHg toxicity to SH-SY5Y neuroblastoma and EA.hy926 vascular endothelial cell lines. In addition, calpain, a Ca2+-dependent protease, is suggested to be associated with the MeHg toxicity. Because MARCKS is known as a substrate of calpain, we studied the relation between calpain activation and cleavage of MARCKS and its role in MeHg toxicity. In SH-SY5Y cells, MeHg decreased cell viability along with increased calcium mobilization, calpain activation, and a decrease in MARCKS amounts. However, pretreatment with calpain inhibitors attenuated the decrease in cell viability and MARCKS amount induced only by 1 μM but not by 3 μM MeHg. In cells with a MARCKS knockdown, calpain inhibitors failed to attenuate the decrease in cell viability caused by MeHg. In EA.hy926 cells, although MeHg caused calcium mobilization and a decrease in MARCKS levels, calpain activation was not observed. These results indicate that the participation of calpain in the regulation of MARCKS amounts is dependent on the cell type and concentration of MeHg. In SH-SY5Y cells, calpain-mediated proteolysis of MARCKS is involved in cytotoxicity induced by a low concentration of MeHg.
To clarify the mechanisms underlying the development of displaced abomasum (DA), we repositioned DA in 12 cows by rolling and observed the course of redisplacement using X-ray fluoroscopy. Displacement of the abomasum was observed to follow three patterns: displacement to the left side of the abdominal cavity through the gap formed at the reticulum, atrium of the rumen and ventral sac of the rumen (n=7); displacement to the left side after cranial movement along the diaphragm (n=3); displacement to the right side after cranial movement along the diaphragm (n=2). These differences were considered to be associated with the rumen volume. To the best of our knowledge, this is the first study reporting the visual observation of the course of DA in dairy cows.
Pancreatic fibrosis develops as the results of the activity of myofibroblasts capable of producing collagens. The myofibroblasts derive from pancreatic interstitial cells, including pancreatic stellate cells (PSCs), which can express glial fibrillary acidic protein (GFAP). First, we investigated the expression patterns of vimentin, desmin, α-smooth muscle actin (α-SMA), Thy-1, and GFAP in the developing rat pancreas (in fetuses at 18 and 20 days, neonates from 1 to 21 days, and adults). Interstitial cells in the developing pancreas expressed vimentin, desmin, GFAP, and Thy-1 at varying degrees; interestingly, the reactivity for desmin and vimentin was the highest in fetuses. GFAP expression was consistent between fetuses and neonates, and Thy-1 reactivity transiently increased after birth; however, α-SMA-positive interstitial cells were rarely seen. Next, we analyzed the immunophenotypical characteristics of myofibroblasts appearing in pancreatic fibrosis in dogs and cats. With increasing fibrotic grade, myofibroblasts showed increased expression of vimentin, desmin, and α-SMA, in addition to increased GFAP expression. Collectively, pancreatic interstitial cells and myofibroblasts may have similar immunophenotypes, and myofibroblasts might originate partly from GFAP-expressing PSCs.
The main objective of this study was to evaluate the effects of transrectal guidance of the ovaries by an assistant on operative time during bovine laparoscopic ovariectomy. Twenty four clinically healthy Holstein dairy cows were divided randomly into two groups. In the transrectal guidance group, an assistant grasped the ovaries via the transrectal route and pulled them to a position where they could be visualized with a camera. On the other hand, the control group was operated without guidance. The time required to remove both ovaries in the guidance group was shorter than that in the control group (P<0.01). We concluded that laparoscopic ovariectomy with transrectal guidance of the ovaries can substantially shorten operative time, thereby greatly contributing to animal welfare and to reducing the burden on the operator.
A 16-year-old castrated male mongrel cat presented with swelling under the left pinna and a 3 -month history of voice change. Laryngeal endoscopy revealed circumferential oedema around the arytenoid cartilages and hypersecretion of saliva. Histopathological examination of the mass around the left ear canal was considered the primary lesion that originated from cutaneous apocrine adenocarcinoma or parotid gland adenocarcinoma, and it metastasized to the larynx, lung and medial retropharyngeal lymph nodes. This report provides new insights into feline laryngeal diseases which could result in laryngeal metastasis with slight mucosal irregularity alone and without obvious radiographic abnormalities. Therefore, histopathological examination should be performed when a cat presents clinical signs such as stridor, dysphonia, or voice change without any mass-forming laryngeal lesion.
Neuroaxonal dystrophy (NAD) is a neurodegenerative disease characterized by severe axonal swelling (spheroids) throughout the nervous system. In dogs, NAD has been reported in several breeds and a missense mutation in PLA2G6 gene has recently been identified in the Papillon dog NAD. Here we performed ultrastructural analysis to clarify the detailed ultrastructural features of the Papillon dog NAD. Dystrophic axons consisted of accumulation of filamentous materials, tubulovesicular structures, and swollen edematous mitochondria with degenerated inner membranes were often observed in the CNS. At axonal terminals, degeneration of presynaptic membrane was also detected. As reported in Pla2g6 knockout mice, mitochondrial and presynaptic degeneration may be related with the pathogenesis of NAD in Papillon dogs.
Although many studies have been conducted worldwide for Equus caballus papillomavirus (EcPV), limited information is available on the virus in Japan. We recently collected one classical viral papillomatosis sample (E150904) from a racing horse in Japan. Papillomavirus infection was confirmed by histopathology, immunohistochemistry and PCR assays, and the sample was diagnosed as epithelial papilloma. Full-length genome of the virus was cloned and sequenced. It was 7,613 bp in length and had the same genome organization with reported EcPV-1. Moreover, phylogenetic analysis based on L1 gene revealed that the infection was caused by a variant of EcPV-1. This is the first report of EcPV infection in Japan, and would further contribute to the molecular epidemiological and pathological studies for EcPV.
A 10-year-old French bulldog presented with an abdominal tumor. Triple-phase helical computed tomography was performed, revealing a hepatic tumor, an enlarged hepatic lymph node, and no masses in other organs. The hepatic tumor demonstrated marked enhancement, similar to that of the aorta in the arterial phase. The tumor had rich vascularization and a hepatic arterio-venous shunt formed between the hepatic artery and middle hepatic vein. The hepatic tumor was surgically removed and histological diagnosis revealed a hepatic carcinoid tumor. During surgery, rapid massive arterial hemorrhage occurred from the site of the incision. The animal died without improvement post-surgery. In the case of an arterio-venous shunt in a hepatic tumor, it is important to be careful to avoid perioperative bleeding.
To establish an accurate, rapid, and a quantifiable method for the detection of Riemerella anatipestifer infection, a widespread infectious disease in birds, we developed a TaqMan-based real-time PCR assay by using DtxR gene-specific primers and a TaqMan probe. The standard curve established with a linear correlation (R2) of 0.998 and efficiency of 99% between the Ct value and the logarithm of the plasmid copy number. The reproducibility and specificity of the real-time PCR assay were confirmed by using plasmids containing DtxR genes or DNAs extracted from well-known bacteria or viruses causing duck diseases. The real-time PCR assay was 100 times more sensitive than the conventional PCR. The results reveal that the established real-time PCR assay might be a useful method for diagnosis and quantitative detection of Riemerella anatipestifer in birds.
This article released online on July 30, 2014 as advance publication has been retracted by the Editorial Board of Journal of Veterinary Medical Science due to a violation of the journal’s “Information for Authors”.