Parafollicular cells (C-cells) exist within the thyroid glands and display different distributions within the glands among mammalian species. In the one-humped camel (Camelus dromedarius), localization of the C-cells remains under debate. We herein investigated appearance of C-cells and the remnants of the ultimobranchial body, origin of C-cells, in the thyroid glands of one-humped camels. Macroscopically, a white mass was present at one-third the length from the cranial end of the thyroid glands where the cranial thyroid artery entered. In addition, large fossae were frequently found adjacent to the white mass. Histologically, the mass was mainly composed of connective tissues, thyroid follicles, and two types of cell clusters: one was composed of cells with clear cytoplasm and the other was composed of non-keratinized epidermoid cells. The mass and the fossae contained p63-positive cells, indicating that they consisted of ultimobranchial body remnants. Calcitonin was expressed in cells with clear cytoplasm, which were localized just beneath the fossae and in the cell clusters of the white mass. C-cells also resided in both subfollicular and interfollicular spaces adjacent to the white mass, but gradually decreased toward the periphery. C-cells tended to display round shapes in the ultimobranchial body remnants and subfollicular spaces, and spindle shapes in interfollicular spaces. In conclusion, we demonstrated that the ultimobranchial body remnants were limited to the region around the entrance of cranial thyroid artery and vein, and C-cells were mainly concentrated within and around the ultimobranchial body remnants.
Live attenuated vaccines are critical in the control of avian infectious bronchitis. It is necessary to know the protection conferred by commonly used commercial live vaccines. In this study, specific pathogen-free chicks were vaccinated with the commercial live vaccines H120, 4/91 and LDT3-A. Blood samples were collected at weekly intervals for the detection of IBV-specific antibodies and quantification of CD4+ and CD8+ T lymphocytes. At 21 days post-inoculation the vaccinated birds were challenged with the IBV prevalent local strains GX-YL5, GX-GL11079 and GX-NN09032, respectively. Trachea and kidney samples were collected at 5 days post-challenge for the detection of the virus. The results showed that the H120 group exhibited medium antibody levels, the lowest percentages of CD4+, CD8+ T lymphocytes and the highest viral loads. The 4/91 group showed the lowest antibody levels, but the highest percentages of CD4+, CD8+ T lymphocytes and the lowest viral loads. The LDT3-A group showed the highest antibody levels, the medium percentages of CD4+, CD8+ T lymphocytes and the medium viral loads. The protection rates of H120, 4/91 and LDT3-A groups were 41.7–58.3%, 75.0–83.7% and 66.7–75.0%, respectively. The present study demonstrated that the vaccines H120, 4/91 and LDT3-A could stimulate the immunized chicks to produce different levels of humoral and cellular immunity to resist the infection of IBV, but couldn’t provide complete protection against the prevalent local strains of IBV in southern China. Also, the vaccine 4/91 offered the best immune protection among the three vaccines.
The aim of this study was to establish a proof-of-concept of protective efficacy of Salmonella-based B. abortus vaccine candidate in Beagles. Group A Beagles (n=10) were subcutaneously (SC) inoculated with S. Typhimurium delivery strain containing pMMP65 (vector to deliver antigens) only as vector control. Group B Beagles (n=10) were SC vaccinated with the mixture of the three Salmonella delivery strains expressing the recombinant B. abortus BCSP31, Outer membrane protein 3b (Omp3b), and superoxide dismutase (SOD) proteins, respectively. No Salmonella delivery strains were isolated from all tissues tested. Serum IgG, interleukin-4, tumor necrosis factor-alpha, and interferon-gamma concentrations were significantly higher in group B than in group A. Following intraconjunctival challenge with B. abortus 544, among 5 group B Beagles, the challenge strain was isolated from mandibular, and retropharyngeal lymph nodes of three Beagles, and no isolates were observed from all tissues of two Beagle. However, the challenge strains were detected from spleen, uterus (except two Beagles), and mandibular, prescapular, retropharyngeal, and superficial inguinal lymph nodes of all group A Beagles. These results suggest that the mixture of three S. Typhimurium delivery strains be a good vaccine candidate against brucellosis by B. abortus in dogs. Further investigations are needed to improve the protective efficacy of the Salmonella-based B. abortus vaccine candidate and explore its practical application in dogs.
The purpose of this study was to determine the relationships among body condition score (BCS), radiography, and computed tomography (CT), and to establish a method for body fat assessment on CT in dogs. Thirty eight Beagles with 2 to 7 BCS were examined. Subcutaneous fat thickness (ST) on radiograph and body area (BA), total fat area (TA), subcutaneous fat area (SA), and visceral fat area (VA) on CT were measured at the level of L3 and L6 vertebra. Ratios of each value to the L6 length were obtained (rST, rTA, rSA, rVA) and the correlations with BCS were estimated. The value of VA/SA, VA/TA, TA/BA, VA/BA, and SA/BA were selected for measuring fat and the correlations with BCS were estimated. The rST, rTA, rSA, and rVA were significantly correlated with BCS, and the rTA and rSA were significantly correlated with rST. At the level of L3, rTA and rVA had stronger relationships with BCS than at L6 while rSA had a higher correlation with BCS at L6. The TA/BA, VA/BA, and SA/BA were significantly correlated with BCS, and the upper limits were 15.11, 6.31, and 8.92%, respectively. Our results showed that CT could be useful to assess body fat and TA/BA, VA/BA, and SA/BA are suitable criteria for measuring fat on CT. In addition, L3 was a more suitable location for evaluating total and visceral fat, and L6 was more suitable for evaluating subcutaneous fat.
A 12-year-old neutered male domestic shorthair cat was presented with a 3-year history of multiple nonpruritic, ulcerated, plaque-like skin lesions but no other clinical signs. A systemic examination revealed mild lymphadenopathy. Histopathologic analysis of the skin lesions revealed multicentric basosquamous carcinoma (BSC). Immunohistochemical analysis, PCR, and sequencing detected Felis catus papillomavirus type 2 (FcaPV-2) within the tumors. As BSC is rare in cats, clinical behavior has not been established. To our knowledge, this is the first case report to demonstrate detection of FcaPV-2 within a BSC in a domestic cat.
In recent years, a relationship between diabetes and neurodegenerative diseases, such as Parkinson’s disease, Alzheimer disease or depression, has been proposed. In this study, pathophysiological changes in the brain, especially in the hippocampus, of male SDT fatty rats with obesity and hyperglycemia were investigated. Brains of SD rats and SDT fatty rats were collected at 32 and 58 weeks of age, and parietal cortical thickness and number of pyramidal cells in the hippocampal cornu ammonis 1 and 3 (CA1 and CA3) regions were measured. At 58 weeks of age, the parietal cortical thickness and number of pyramidal cells in the hippocampal CA1 and CA3 regions were lower in SDT fatty rats than in age-matched SD rats. Measurements of mRNA in rat brains at 58 weeks of age showed that the expression of genes related to inflammatory responses (S100a9, TNFα, NF-κB) was elevated in SDT fatty rats. From the aforementioned results, changes suggestive of brain atrophy and impairment in cognitive function were observed in male SDT fatty rat brains.
The effect of riboflavin supplement in Eimeria tenella-infected chickens was evaluated. Mortality, fecal consistency, and oocysts per gram of feces were monitored for groups of E. tenella-infected chickens administered a basal diet supplemented with either riboflavin, the anti-coccidial drug amprolium, or with both compounds. The number of oocysts shed per chicken in the riboflavin-treated group was significantly higher than the positive non-treated control group. No significant difference in oocyst number between the amprolium-treated group and riboflavin plus amprolium-treated group was observed. Thus, the addition of 0.8 g/kg of riboflavin to basal diet can increase oocyst number in E. tenella-infected chicken, but has no effect on the efficacy of amprolium.
Bats are potential reservoirs of Cryptosporidium and Eimeria. The genus Cryptosporidium infects various vertebrates and causes a diarrheal disease known as cryptosporidiosis. Many epidemiological studies in wild animals have been performed; however, most of them relied on only PCR-based detection because of the difficulty of performing pathological analyses. Accordingly, the natural host and pathogenicity of Cryptosporidium bat genotypes remain unclear. In this study, we captured Eptesicus nilssonii (Northern bats) in Hokkaido, Japan. Of the three intestinal samples obtained, two were positive for Cryptosporidium spp. and one was positive for Eimeria spp. The corresponding microorganisms were also confirmed histopathologically. We detected the novel Cryptosporidium bat genotype XII and Eimeria rioarribaensis in bat intestine.
Polyomaviruses (PyVs) infect a wide range of animals and provoke wasting diseases, particularly in immunosuppressed hosts. Recently, a novel Rattus norvegicus polyomavirus 2 (RatPyV2) has been identified in a colony of X-linked severe combined immunodeficiency (X-SCID) rats in the United States. Here, we describe the first report of the RatPyV2 infection in an X-SCID rat colony in Japan. The affected rats exhibited adult-onset wasting. Histologically, we observed large basophilic intranuclear inclusion bodies within the hyperplastic or dysplastic epithelial cells in the salivary glands, Harderian glands, extraorbital lacrimal glands, and in respiratory and reproductive tissues. Among these organs, the parotid salivary, Harderian, and extraorbital lacrimal glands were most obviously affected. In particular, the parotid salivary glands were the most severely and diffusely affected and atrophic lesions were prominent even at 1 month of age, which suggested that the parotid salivary glands would be highly susceptible to RatPyV2 in X-SCID rats. RatPyV2 inclusion bodies were also detected in the tail of the epididymis and deferent duct. Such reproductive lesions developed significantly in the later stage of breeding age, and therefore may be associated with the reduced fecundity observed in the infected X-SCID rats. We also established a simple, rapid, and non-invasive diagnostic method based on the Amp-FTA method, using buccal swabs for the detection of RatPyV2 in immunodeficient rats. Our findings contribute to the early detection and diagnosis of RatPyV2 infections.
ML204, a potent transient receptor potential canonical 4 (TRPC4) channel blocker, is often used to elucidate the involvement of TRPC4 channels in receptor-operated signaling processes in visceral smooth muscles. In the present study, we investigated the possible antagonistic actions of ML204 on M2 and M3 muscarinic receptors, which mediate contractions in mouse ileal and detrusor smooth muscles. In ileal and detrusor smooth muscle preparations, ML204 (3 or 10 µM) significantly inhibited electrical field stimulation (EFS)-evoked cholinergic contractions. However, it did not significantly inhibit high K+-induced and EFS-evoked non-cholinergic contractions in the ileal preparations. When the muscarinic agonist, carbachol was cumulatively applied, ML204 (1, 3 and 10 µM) caused a rightward parallel shift of the concentration-response curves of carbachol. Additionally, ML204 (1, 3 and 10 µM) inhibited carbachol-induced negative chronotropic response in atrial preparations, which is mediated by M2 muscarinic receptors. Furthermore, ML204 significantly inhibited the contractions evoked by carbachol-induced intracellular Ca2+ release, which is mediated by M3 muscarinic receptors. These results suggested that ML204 might exhibit antagonistic actions on M2 and M3 muscarinic receptors; in addition, the inhibitory effects of ML204 against EFS-induced cholinergic contractions might be attributed to this receptor antagonism rather than inhibition of TRPC4 channel activity. Therefore, these effects should be considered when ML204 is used as a TRPC4 channel blocker.
We succeeded in the simultaneous isolation of Brachyspira (B.) aalborgi and B. pilosicoli from a patient with ulcerative colitis. B. pilosicoli grew quickly and formed colonies within 7 days, while the growth of B. aalborgi was very slow and took over 21 days. Simultaneous isolation of B. pilosicoli and B. aalborgi from a common specimen is generally recognized to be difficult, mainly due to differences in their growth requirements and the growth rates. However, we succeeded in isolating both species from a patient with ulcerative colitis and this is first evidence. The present results suggest that ulcerative colitis may be caused by simultaneous infection with B. pilosicoli and B. aalborgi.
Robenacoxib is a novel nonsteroidal anti-inflammatory drug approved for dogs. The present study aimed to evaluate influences of sevoflurane anesthesia on the distribution of robenacoxib in dogs. Ten healthy beagle dogs (1 to 11 years old, 9.3 to 14.3 kg body weight, 6 males and 4 females) were subcutaneously administered robenacoxib (2 mg/kg) under conscious condition or sevoflurane anesthesia inhaled a 1.3-fold predetermined individual minimum alveolar concentration of sevoflurane at a 28-day interval. The dogs under sevoflurane anesthesia were also mechanically ventilated and received fluid-therapy. On each occasion, serum samples were collected from the dogs before and at 5, 15, 30, 60, 120, 180, and 240 min after the robenacoxib administration. Serum robenacoxib concentration was measured by a liquid chromatography-tandem mass spectrometry. Maximum serum concentration of robenacoxib (Cmax) was 2.2 µg/ml [range: 1.2–4.6] (median [range: minimum-maximum]) and time of Cmax (Tmax) was 90 min [range: 60–120] in the conscious dogs. In the sevoflurane-anesthetized dogs, the Cmax significantly declined (1.3 µg/ml [range: 0.8–1.4], P=0.008) and Tmax was delayed (120 min [range: 120–240], P=0.018) compared with those in the conscious dogs. The serum robenacoxib concentration at 240 min (C240) decreased to 0.5 µg/ml [range: 0.2–0.9] in the conscious dogs, while it remained higher in the sevoflurane-anesthetized dogs (1.0 µg/ml [range: 0.3–1.4], P=0.011). In conclusion, the anesthetic procedure with sevoflurane, mechanically ventilated, and received fluid-therapy might affect the pharmacokinetics of subcutaneously administered robenacoxib in dogs.
The relationships between propofol plasma concentrations and the pharmacodynamic endpoints may differ according to a type of airway device. To clarify these relationships in different airway devices would be useful to avoid the complication such as apnea and intraoperative awareness. The aim of this study was to investigate the influence of difference of airway device on propofol requirement during maintenance of anesthesia in dogs. We compared the influence of airway devices on the plasma propofol concentrations for apnea, response to mechanical ventilation, and response to airway device between endotracheal tube (ETT) and supraglottic airway device (SGAD) in Beagles. The pharmacodynamic effects were repeatedly assessed at varying propofol concentrations. The plasma concentrations (mean ± SD) of propofol in the ETT and SGAD groups were 10.2 ± 1.8 and 10.9 ± 2.4 µg/ml for apnea (P=0.438), 7.9 ± 1.2 and 7.4 ± 1.5 µg/ml for response to mechanical ventilation (P=0.268), and 5.2 ± 0.7 and 5.4 ± 1.5 µg/ml for response to airway device (P=0.580), respectively. Required propofol concentration during maintenance of anesthesia may be similar between ETT and SGAD. Without moderate to strong stimuli such as airway device insertion or painful stimulation during surgery, the type of airway device may have little impact on required propofol concentration during maintenance of anesthesia in dogs.
A 10-year-old cat presented for evaluation with a 1-month history of salivation and oral bleeding. A right mandibular mass was palpated and computed tomography examination revealed entire bone proliferation. Mandibular bone biopsy was performed, and histopathological diagnosis was vascular hamartoma. The cat suddenly died on day 140.
Rabies virus (RABV), canine distemper virus (CDV), canine parvovirus type-2 (CPV-2), and canine influenza A virus (CIV) are important contagious pathogens in canine populations. To assess post-vaccination immunity against RABV, CDV and CPV-2, and serological evidence of exposure to influenza A virus in military working dogs (MWDs) in Korea, we tested blood samples of 78 MWDs by fluorescent antibody virus neutralization (FAVN) for RABV, and by commercially available enzyme-linked immunosorbent assay (ELISA) for CDV, CPV-2, and CIV. Korean MWDs had high antibody-positive rates against RABV (97.4%, ≥0.5 IU/ml), CDV (94.8%), and CPV (100%). All dogs tested seronegative (0/78; 0%) for influenza A virus. Two 1-year-old dogs stationed in known rabies outbreak areas (Gangwon and Gyeonggi) exhibited VNA titers below the protective level (0.06 and 0.29 IU/ml, respectively). The breed and sex of MWDs were not significantly associated with antibody titers for RABV, CDV, or CPV; however, age was significantly associated with CPV antibody titers, while region of residence was associated with CDV antibody titer. Taken together, the data presented here provide important insights necessary for post-vaccination management and control of infectious diseases in MWDs.
Zoonoses are frequently reported, and outbreaks of the highly pathogenic influenza virus, severe acute respiratory syndrome, and Middle East respiratory syndrome have occurred recently, in Africa, the Middle East, and Southeast Asia. Sterilization using a chemical reactor with plasma assisted catalytic technology (PACT) was investigated. Tests were carried out on the feline calicivirus (FCV) vaccine strain F9, which is a surrogate of airborne pathogen human norovirus. Results showed that the PACT device could inactivate FCV, which passed through the plasma chamber. Sterilization rate may be more than 99.99% (below the detection limit). These results indicate that PACT may be an effective mean to inactivate many viruses, including human norovirus, and potentially other airborne, infectious microorganisms.
The objective of this study was to compare the efficacy of a commercial porcine reproductive and respiratory syndrome (PRRS) subunit vaccine and a prototype PRRS II subunit vaccine against a highly pathogenic PRRS virus (HP-PRRSV) in pigs. Both vaccines were administered intramuscularly in 2 doses at 21 and 42 days of age, and the pigs were challenged intranasally with HP-PRRSV at 63 days of age. Pigs vaccinated with the prototype PRRS II subunit vaccine had significantly higher anti-PRRSV antibody titers, a greater number of interferon-γ-secreting cells, and a greater reduction in lung lesion scores compared to pigs vaccinated with the commercial PRRS subunit vaccine. Therefore, the commercial PRRS subunit and prototype PRRS II subunit vaccines are efficacious against HP-PRRSV.
Porcine circovirus associated diseases (PCVAD) have multiple manifestations that have been attributed to porcine circovirus type 2 (PCV2). Recently, a novel porcine circovirus, PCV type 3 (PCV3), was identified in pigs with systemic inflammation of unknown etiology. In this study, we tried to detect the PCV3 genome in tissue samples collected from Japanese pig herds in 2016. The PCV3 genome was detected by PCR in 7 of 73 samples. The homology between each Japanese strain was 99.5% for the full-length sequence and 98.9 to 99.2% for the open reading frame 2. These results suggest that PCV3 has already invaded Japanese pig farms.
Long-term monitoring of circulating progesterone levels in three captive female false killer whales, Pseudorca crassidens, was conducted to characterize their reproductive events and to reveal the relationship between their estrous cycles or pregnancies and peripheral white blood cell (WBC) counts. Blood samples were collected at 2–3-day intervals or on a weekly-to-monthly basis for up to 10 years, from 2006 to 2017. In two mature females (initial body lengths of 4.22 and 4.07 m), some cyclic progesterone elevations were detected during the study period; the estimated mean (± SE) estrous cycle length was 40.5 ± 0.7 days (n=12). The seasonality of ovulation, estimated from the elevation of progesterone levels, varied among individuals or years, and ovulation did not occur every year. The third female (3.26 m) showed progesterone elevations, despite irregular cycles after sexual maturity, and became pregnant. The progesterone levels during pregnancy ranged from 7.3 to 42.2 ng/ml, and the gestation period lasted for 14 months until parturition. The mean WBC counts during estrous cycles were the lowest before the progesterone levels began to increase and then gradually increased toward the luteal phase. The WBC counts were significantly higher during pregnancy than before and were particularly high in early pregnancy. To the best of our knowledge, this is the first report of the relationship between the estrous cycle or pregnancy and WBC counts in cetaceans.
The purpose of this study was to assess tick-borne pathogenic infections in 42 wild Korean water deer (KWD) and 26 farmed elk in the Gyeongbuk and Gangwon Provinces of Korea. Among the 42 wild KWD tested, the eighteen (42.9%) and five (11.9%) samples tested positive for Anaplasma phagocytophilum and A. bovis, respectively, by PCR and DNA sequencing. All positive samples were only from wild KWD. All samples were negative for other tick-borne pathogens tested. Detected 16S rRNA sequences of A. phagocytophilum and A. bovis showed 98.6–99.8% and 94.4–100% identity to those of sequences in GenBank, respectively. Because few studies have examined tick-borne pathogens in wild animals, appropriate control programs and studies are needed to prevent pathogen transmission.