Monophasic variants of Salmonella enterica serovar Typhimurium isolated in Thailand and Japan were characterized to elucidate the genetic basis of the monophasic phenotype, genetic relatedness, and antimicrobial resistance. A total of 20 Salmonella isolates agglutinated with anti-O4 and anti-H:i serum and not agglutinated with either anti-H:1 or anti-H:2 serum were identified as monophasic variants of Salmonella serovar Typhimurium because they harbored IS200, specific to this serovar, and lacked the fljB gene. An allele-specific PCR-based genotyping method that detects a clade-specific single nucleotide polymorphism indicated that seven swine isolates and one human isolate from Thailand were grouped into clade 1; five isolates from layer chicken houses and layer chicken feces from Japan were grouped into clade 8, together with two Salmonella serovar Typhimurium isolates from chicken houses in Japan; and five isolates from swine feces from Thailand and two isolates from layer chicken feces from Japan were grouped into clade 9. Multilocus sequencing typing demonstrated that sequence type (ST) 34 isolates were solely grouped into clade 9. Clade 1 and 8 isolates were assigned as ST19. Pulsed-field gel electrophoresis revealed multiple types within each of the clades. The presence of antimicrobial resistance genes and plasmid replicon type, of the clade 1 and 9 isolates were comparable to those reported for epidemic strains of monophasic variants. Our results suggest that monitoring monophasic variants of serovar Typhimurium is important for understanding of the spread of these variants in Thailand and Japan.
The control of inflammation, which arises from complex biological responses to harmful stimuli, is an important determinant of both clinical outcomes and patient comfort. However, the side effects of many current therapies such as non-steroidal anti-inflammatory drugs mean that new safe treatments are required. We previously reported that 12.5 µg/ml hydroxytyrosol (HT) suppressed gene expression of the inducible nitric oxide (NO) synthase (iNOS) isoform and NO production, in mouse peritoneal macrophages treated with lipopolysaccharide (LPS), where nuclear factor-κB (NF-κB) gene expression was not altered. The present study evaluated the anti-inflammatory effects of various concentrations of HT in LPS-induced RAW264.7 mouse macrophages. HT suppressed NF-κB signaling and downregulated LPS-mediated expression of iNOS, cyclooxygenase-2, tumor necrosis factor alpha, and interleukin-1β at 12.5 µg/ml, resulting in reduced production of NO and prostaglandin E2. At lower concentrations, HT seemed to act via another signaling pathway to regulate the inflammatory response. In contrast, HT did not suppress LPS-induced expression of phosphorylated p44/42 mitogen-activated protein kinase. This study showed that HT had anti-inflammatory effects on LPS-stimulated RAW264.7 cells. HT is already available as a nutritional supplement and no toxic effects have been reported. Hence, HT represents a potential novel anti-inflammatory agent.
This study assessed the advantages of dextrose and amino acid mixture solution as parenteral nutrition (PN) therapy for diarrheic calves. Thirty diarrheic calves were randomly assigned to receive PN (PN group, n=15) or only dextrose solution (Dex group, n=15). The treatment period for the PN group (4.0 days; min-max, 2–10 days) was significantly shorter than that for the Dex group (6.0 days; min-max, 3–21 days) (P<0.01). The PN therapy tended to improve plasma diamine oxidase activity compared with traditional therapy. One potential association between PN therapy and shortened treatment period may be the repair of damaged intestinal villi. Although our proposal has limitations, PN therapy suggested the potential for new treatment of diarrheic calves.
An 11-year-old cat presented with nasal discharge and lacrimation and was diagnosed with nasal lymphoma. Although the cat showed favorable progression after undergoing chemotherapy, CT imaging demonstrated enlarged pulmonary nodules caused by Toxoplasma gondii. Following the cessation of chemotherapy, the cat was prescribed clindamycin hydrochloride for toxoplasmosis treatment; however, the cat developed kidney lymphoma and died. No T. gondii organisms were observed in the whole body necropsy specimens. It is known that immunocompromised human patients, including those who undergo chemotherapy, are considered at risk for toxoplasmosis. However, the risk of developing toxoplasmosis in cats undergoing chemotherapy is currently unknown. Findings from this case report suggest that cats with chemotherapy-resistant pulmonary masses might have a T. gondii infection rather than metastatic disease.
α-defensin is a potent antimicrobial peptide secreted from intestinal mucosal epithelial cells, such as Paneth cells, and affects not only bacteria but also parasites and fungi. Recently, human eosinophils have also been shown to produce α-defensin, but no studies have been done on other animals. In this study, we attempted to detect α-defensin protein in mouse eosinophils infiltrating the intestinal mucosa during a helminth infection using Zamboni fixation and immunohistochemistry. Most of the eosinophils infiltrating the intestinal mucosa during helminth infection were positive for α-defensin. The expression level of α-defensin mRNA was 50 fold that in the control. Meanwhile, the number of Paneth cells was doubled, and their α-defensin fluorescence intensity was increased. These results suggested that eosinophils are also important producers of α-defensin, such as Paneth cells in mice, and that α-defensin produced from eosinophils might be involved in defensive mechanisms against helminths. Moreover, the experimental system used in this study is a good model to study the generation of α-defensin by eosinophils.
The present study describes the association between inflammatory cell types and feline infectious peritonitis virus (FIPV) antigen in the brain of 4 cats diagnosed as feline infectious peritonitis (FIP). Immunohistochemically, FIPV antigens were detected in the inflammatory foci of the leptomeninges, choroid plexus and ventricles in 3 of the 4 cats. In 3 cases, inflammatory foci mainly consisted of CD204- and Iba1-positive macrophages, and the FIPV antigens were found in the macrophages. In the other case which was negative for FIPV antigen, severe inflammation predominantly consisting of CD20-positive B lymphocytes was observed in the leptomeninges and subventricles, accompanied with diffuse proliferation of gemistocytic astrocytes. The difference in histopathology may reflect the inflammatory process or the strain variation of FIP virus.
Recently, we reported that follicle-sinus complexes (FSCs) in the muzzle skin are useful for postmortem diagnosis of rabid dogs. Here, we compared the sensitivity and specificity of detecting the viral antigen in the brain and FSCs of 226 suspected rabid dogs, and assessed whether the FSC harbored the virus genome and particles. The viral antigen was detected in 211 of 226 samples with 100% sensitivity and specificity. Viral RNA and particles were observed in the cytoplasm of Merkel cells (MCs). These results suggest that MCs are targets of virus infection and FSCs are useful material for diagnosing rabies.
The purpose of this study was to evaluate the bioequivalence of 5% ceftiofur hydrochloride sterile suspension in two formulations, a test formulation (Saifukang 5% CEF, Hvsen) and a reference formulation (Excenel®RTU 5% CEF, Pfizer). Twenty-four healthy pigs were assigned to a two-period, two-treatment crossover parallel trial, and both formulations were administered at a single intramuscular dose of 5 mg/kg weight, with a 7-day washout period. Blood samples were collected consecutively for up to 144 hr after administration. The concentrations of ceftiofur- and desfuroylceftiofur-related metabolites in the plasma were determined by high-performance liquid chromatography. In addition, the major pharmacokinetic parameters (Cmax, AUC0-t and AUC0-∞) were computed and compared via analysis of variance, with 90% confidence intervals. Bioequivalence evaluation of Tmax was statistically analyzed with the nonparametric test. The comparison values between test and reference formulation for AUC0-t, AUC0-∞, Cmax, and Tmax were 376.7 ± 75.3 µg·hr/ml, 390.5 ± 78.6 µg·hr/ml, 385.9 ± 79.2 µg·hr/ml, 402.7 ± 80.4 µg·hr/ml, 34.6 ± 5.5 µg/ml, 36.1 ± 6.2 µg/ml, 1.27 ± 0.18 hr, and 1.26 ± 0.21 hr, respectively, and we observed no significant differences between the two formulations. The 90% CI values were within the recommended range of 80–125% (P>0.05), and the relative bioavailability of the test product was 96.47 ± 10.92% according to AUC0-t values. Based on our results, the two formulations exhibit comparable pharmacokinetic profiles, and the test product is bioequivalent to the reference formulation.
The aim of the study was to evaluate effects of fisetin and probiotic on erythrocyte osmotic fragility (EOF), malondialdehyde (MDA) and superoxide dismutase (SOD) in broiler chickens exposed to heat stress. Sixty day-old broilers were divided into: Group I (control) given distilled water; Group II, fisetin (5 mg/kg); Group III, probiotic Saccharomyces cerevisiae (4.125 × 106 cfu/100 ml); and Group IV, fisetin (5 mg/kg) + probiotic (4.125 × 106 cfu/100 ml) orally for 7 days. Blood samples collected from 42-day-old birds were evaluated for EOF, serum MDA concentration and SOD activity. Percentage EOF at 0.5% NaCl was lower (P<0.05) in fisetin, probiotic and fisetin + probiotic groups (34.26 ± 0.98%, 35.65 ± 0.81% and 34.25 ± 1.98%, respectively) than in controls (48.42 ± 0.40%). The MDA concentrations in broiler chickens administered with fisetin (14.37 ± 1.15 nmol/l), probiotic (5.66 ± 1.06 nmol/l) and fisetin + probiotic (4.136 ± 0.58 nmol/l) were lower (P<0.05) than in controls (22.64 ± 2.95 nmol/l). Activities of SOD were higher (P<0.05) in fisetin, probiotic and fisetin + probiotic broiler chickens (6.34 ± 0.24 IU/l, 5.67 ± 0.09 IU/l and 5.93 ± 0.13 IU/l, respectively) than in controls (5.37 ± 0.09 IU/l). Fisetin + probiotic ameliorated oxidative stress changes in broiler chickens better than fisetin or probiotic alone. In conclusion, administration of fisetin or probiotic and, especially their combination, decreased EOF, lipoperoxidation and increased superoxide dismutase activity in broiler chickens exposed to heat stress.
In Japan the highest use of veterinary antimicrobials is in pig production. To obtain useful information to achieve the best approach to reducing this use, we analyzed the association between the level of on-farm biosecurity and animal welfare with the level of antimicrobial use as recorded on prescriptions on 38 pig farms under contract to veterinarians of the Japanese Association of Swine Veterinarians. To determine the level of welfare we recorded the risk of pre- and post-weaning deaths and the floor space available per fattening pig (m2/head). Multivariable linear regression analysis was performed, using biosecurity scores and animal welfare indicators as independent variables and the amount of antimicrobial usage as dependent variables. The results showed that the higher scores for the site condition (location) and external biosecurity scores of the sub-categories ‘farm contractors’ were strongly associated with the lower use of oral antimicrobials (P<0.05). This suggests that in order to reduce the usage of antimicrobials for herd treatments, farmers should consider the location when building a new farm or pig house and strengthen the entrance requirements for high risk visitors. Regression analysis for the respective antimicrobials showed that the site condition, the biosecurity scores of the sub-categories ‘farm contractors’, ‘pen layouts’ (e.g. independence of pens and sites), ‘pig flows’ (e.g. the completeness of all-in/ all-out system) and an animal welfare indicator (i.e. post-weaning mortality risk) were significantly associated with the use of one or more antimicrobials (P<0.05).
A 4-year-old, male Bernese mountain dog was evaluated for a 1-year history of right hemiparesis. Computed tomography revealed a large hypoattenuating mass severely deforming the C5 vertebral arch, invading the C6 spinal canal, and causing spinal cord compression. The signal characteristics of magnetic resonance imaging indicated a lesion composed of adipose tissue. The mass was removed via right hemilaminectomy, and histopathological examination confirmed it was an infiltrative lipoma. The compressive lesion remained unresolved, so the dog underwent a second operation, after which he regained some ambulatory function. Although postoperative adjunctive radiation therapy was performed, the dog died 201 days after the first operation.
The objective of this study was to evaluate the effects of post artificial insemination (AI) treatment with intravaginal progesterone device (P4 device) on conception rate, synchronization of returning estrus and plasma P4 concentration in Japanese Black cows. Nineteen cows were treated with DIB (1.0 g P4) from Day 12 to 19 (Day 0=day of the first AI), 27 cows were treated with a CIDR (1.9 g P4) from Day 12 to 19, and 33 cows were not treated after the first AI (control). Estrous behavior was daily examined between Day 20 and 25, and cows returning to estrus were inseminated (the second AI). On Day 19, plasma P4 concentration was not different among DIB, CIDR and control groups. There was no significant difference in conception rate after the first AI among three groups (DIB: 63.2%, CIDR: 66.7% and control: 72.7%). In non-pregnant cows, there was no significant difference in the proportion of cows showed returning estrus between Day 20 and 25 (DIB: 57.1%, CIDR: 22.2% and control: 44.4%), and day of returning estrus was not synchronized. The overall conception rate after the first and second AI was not different among the groups. In conclusion, post-AI treatment with intravaginal devices containing 1.0 and 1.9 g P4 from Day 12 to 19 neither increased plasma P4 concentration nor improved fertility and synchronization of the returning estrus in Japanese Black cows.
Stem cell therapy has been proposed to restore the function and structure of injured tissues. In the present study, we investigated the ability of human endothelial progenitor cells (hEPCs) to attenuate ovarian aging and dysfunction. Female ICR mice aged 4 and 6 months were injected with cultured hEPCs. Cultured hEPCs were injected intravenously twice with 5 × 104 cells with a 4 day interval. After pregnant mare serum gonadotropin and human chorionic gonadotropin stimulation, oocytes and ovaries of aged mice were collected, cumulus-free oocytes were activated by SrCl2 and gene expression levels related to inflammation, apoptosis, follicle development and endoplasmic reticulum (ER) stress in ovaries were compared. Administration of hEPCs attenuated the level of inflammatory cytokines and adverse apoptotic factor, as well as reducing ER stress in the ovaries. Increased cleavage and blastocyst formation rates and cell numbers in blastocysts from hEPCs-treated aged mice vs. same aged control mice demonstrated a protective function of hEPCs against reproductive aging. Based on these data, we suggest that treatment with hEPCs attenuates reproductive aging and dysfunction potentially via regulation of inflammation, apoptosis and ER stress.
Plasma luteinizing hormone (LH) and testosterone concentrations were examined in Japanese Black beef bulls with normal and abnormal semen in response to gonadotropin releasing hormone (GnRH) challenge at the start (10 months) and completion (20 months) of puberty. Bulls with normal semen had higher testosterone concentrations after GnRH treatment at 20 months than they did at 10 months, while LH concentrations did not differ between the two age groups. LH and testosterone concentrations were not different between bulls with normal and abnormal semen at 20 months. Thus, testosterone secretions in response to the GnRH challenge were higher for bulls with normal semen at pubertal completion compared to bulls at the start of puberty, but responsiveness of LH to GnRH and of testosterone to the LH increment was not altered in bulls with abnormal semen.
Muscovy duck parvovirus (MDPV) and goose parvovirus (GPV) have both been found to cause high mortality and morbidity in Muscovy ducklings. Specific detection is often rife with false positives due to high identity at the genomic nucleotide level and antigenic similarity between MDPVs and GPVs. In this study, significantly variable regions were found, via non-structural (NS) comparison, between MDPV and GPV NS genes; however, NS genes were conserved within the MDPV and GPV groups. A polymerase chain reaction (PCR) assay for detecting and differentiating MDPVs and GPVs was developed with more specificity based on the NS gene characterization. The assay detected as low as 103 DNA copies of both the MDPV and GPV strains, along with 549 separate base pairs (bp). No bands of the same size from other duck pathogens, including duck circovirus, duck enteritis virus, egg drop syndrome virus, duck-origin goose hemorrhagic polyomavirus, Escherichia coli, Salmonella, Riemerella anatipestifer and Pasteurella multocida were amplified. This indicates that this method for performing PCR provides a useful and reliable alternative tool for more precise differentiation of MDPV and GPV infection in clinical samples.
The Sunda pangolin (Manis javanica) population in Southeast Asia faces threats such as poaching and deforestation. Health assessments of rescued individuals including physical examination and blood work are crucial for clinicians to determine the health status of these animals. The establishments of reference intervals of hematology and serum biochemistry are important for identifying clinical abnormalities. The objectives of our study were to establish blood reference intervals for Sunda pangolins, to determine if there are age and sex related differences in hematology and serum biochemistry, and to compare our results with those of a previous study on confiscated Sunda pangolins in Thailand. Fifty-eight Sunda pangolins were rescued between January 2011 and December 2015. The hematology and serum biochemistry results of 51 clinically normal Sunda pangolins were selected for the establishment of the blood reference intervals. No sex related differences were noted in this study. Age-related differences were observed, in which adult Sunda pangolins had a significantly higher mean corpuscular volume than juveniles, and juvenile Sunda pangolins had significantly higher red blood cell counts and hemoglobin levels than those of the adults (P<0.05). Age-related differences were also noted in several serum biochemistry parameters: alkaline phosphatase (ALP) was significantly higher in juveniles, and total protein was significantly higher in adult Sunda pangolins. Compared to a previous study the white blood cell counts, neutrophil counts, and ALP were higher, and the lymphocyte counts were lower in the present study.
Asian and African elephants are frequently afflicted by foot disorders that can be very challenging to manage even with aggressive therapy. Such conditions may have indirect life-threatening effects. Mohs’ paste (zinc chloride based escharotic agent) was used to treat a female Indian elephant (Elephas maximus indicus) aged 39 years with foot disorder at Kanazawa Zoological Gardens. Degenerated hyperplastic tissue was observed inside the hoofs of digits 2 and 5. Mohs’ paste was applied on the lesions, which coagulated the hyperplastic tissue and restrained its proliferation. Subsequently, the hyperplastic tissue could be trimmed with little pain, and the disorder became manageable. Mohs’ paste treatment was effective and is expected to be an alternative treatment for hoof disorder.
In this study, we aimed to report anthrax cases in two pumas, brought to the Pathology Department, Faculty of Veterinary Medicine, Erciyes University for suspected poisoning upon their sudden death at the Kayseri Zoo, in Turkey. In the necropsy, enlargement and malacia were observed in the spleens. The cut surfaces of the spleens were in extreme red-blackish color. Bacillus anthracis was isolated as a pure culture from both samples which belong to dead pumas. B. anthracis isolates had pXO1 and pXO2 plasmids. Both isolates were found to be sensitive to eight antibacterials tested. This study demonstrates that feeding of the wild carnivorous kept in any zoo with the appropriate meats which belongs to healthy animals is extremely important.
Although Moraxella lacunata causes conjunctivitis, keratitis, endocarditis, and otolaryngitis in humans, its infection is rare in animals. We report three cases of asymptomatic pulmonary abscesses caused by M. lacunata in zoo herbivores, including two elks (Cervus canadensis) and a common eland (Taurotragus oryx). In all cases, macroscopic findings included coalescence of lung lobes and severe pulmonary abscesses filled with cheese-like materials in cysts. Microscopic findings included pneumonia characterized by marked fibrin exudates in alveolar spaces and infiltration of inflammatory cells. M. lacunata was identified in bacterial cultures from pulmonary abscesses using biochemical API 20NE system. M. lacunata is rarely isolated from zoo animals; however, herein, we describe the first report of pulmonary abscesses caused by M. lacunata infection.