The bacterium Haemophilus parasuis (H. parasuis) is the primary cause of Glässer’s disease. Currently, there are no effective vaccines that can confer protection against all H. parasuis serovars. Therefore, the present study aimed to investigate the effect of tea polyphenols on growth, expression of virulence-related factors, and biofilm formation of H. parasuis, as well as to evaluate their protective effects against H. parasuis challenge. Our findings demonstrated that tea polyphenols can inhibit H. parasuis growth in a dose-dependent manner and attenuate the biofilm formation of H. parasuis. In addition, tea polyphenols exerted inhibitory effects on the expression of H. parasuis virulence-related factors. Moreover, tea polyphenols could confer protection against a lethal dose of H. parasuis and can reduce pathological tissue damage induced by H. parasuis. In summary, our findings demonstrated the promising use of tea polyphenols as a novel treatment for H. parasuis infection in pigs.
Streptococcus parasuis has recently been removed taxonomically from Streptococcus suis, a zoonotic pathogen. S. parasuis has been detected in healthy pigs and in diseased pigs, which suggests that S. parasuis is involved in the normal microbiota of pigs and has potential pathogenicity. However, the pathogenicity of S. parasuis in pigs is unclear because of the lack of appropriate detection methods that discriminate S. parasuis from S. suis. In this study, we developed a PCR method that is specific for S. parasuis. The detection limit of the PCR was 350 CFU per reaction. Bacteria isolated from the saliva of eight pigs were collected and examined by PCR. Sixty-four isolates positive for PCR were obtained from the samples of all pigs. Thirteen of the 64 isolates were genetically confirmed as S. parasuis, and biologically and biochemically had nearly the same features of known S. parasuis strains, which suggested that strains positive for PCR were S. parasuis. Among the 64 isolates, 28 isolates were serotypes 20, 22, or 26 in the S. suis serotyping scheme. The remaining 36 isolates were untypeable, which suggested the presence of novel serotypes or a capsule-negative form. Therefore, the PCR method described in this study is a useful tool for identifying S. parasuis, and can be used in etiological studies on this bacterium.
Wild animals typically exhibit defensive behaviors in response to a wider range and/or a weaker intensity of stimuli compared with domestic animals. However, little is known about the neural mechanisms underlying “wariness” in wild animals. Wild rats are one of the most accessible wild animals for experimental research. Laboratory rats are a domesticated form of wild rat, belonging to the same species, and are therefore considered suitable control animals for wild rats. Based on these factors, we analyzed structural differences in the brain between wild and laboratory rats to elucidate the neural mechanisms underlying wariness. We examined wild rats trapped in Tokyo, and weight-matched laboratory rats. We then prepared brain sections and compared the basolateral complex of the amygdala (BLA), the bed nucleus of the stria terminalis (BNST), the main olfactory bulb and the accessory olfactory bulb. The results revealed that wild rats exhibited larger BLA, BNST and caudal part of the accessory olfactory bulb compared with laboratory rats. These results suggest that the BLA, BNST, and vomeronasal system potentially contribute to wariness in wild rats.
The aim of the present study was to examine the effect of 28 days of dietary difructose anhydride (DFA) III supplementation on calcium (Ca) metabolism in late-lactation dairy cows. Twenty-four multiparous pregnant Holstein cows were divided into two groups. The DFA group was fed total mixed ration (TMR) supplemented with 40 g of DFA III, and the control group was fed TMR only. The replenishment of bone Ca reserves was evaluated by measuring bone mineral density (BMD) and blood biochemical bone markers. Serum Ca concentrations, urinary Ca-to-creatinine (Cre) (Ca/Cre) ratios, and milk Ca concentrations were also analyzed. The BMD of the 4th caudal vertebra in the DFA group was higher than in the control group on day 28. With respect to bone markers, the ratios of undercarboxylated osteocalcin (ucOC) to osteocalcin (OC) in the DFA group were significantly lower than those in the control group on days 21 and 28. Milk Ca concentrations in the DFA group were also higher than those in the control group on days 14, 21, and 28, whereas serum Ca concentrations and urinary Ca/Cre ratios were unchanged in both groups. These results suggest that dietary supplementation with DFA III increased BMD and decreased serum ucOC/OC ratios in late-lactation dairy cows; this indicates that the replenishment of bone Ca reserves may be enhanced by dietary DFA III supplementation.
Sirtuin-1 (SIRT1) is a nicotinamide adenine dinucleotide (NAD+)-dependent histone deacetylase with a large number of protein substrates. It has attracted a lot of attention in association with extending lifespan. The objective of this study was to enable the evaluation of SIRT1 expression in peripheral blood mononuclear cells (PBMCs) from dogs by flow cytometry. Three transcript variants were amplified from PBMCs by reverse transcription PCR and the nucleotide sequences were analyzed. On the basis of deduced amino acid sequence, a monoclonal antibody against human SIRT1, 1F3, was selected to detect canine SIRT1. Canine SIRT1 in peripheral blood mononuclear cells was successfully detected by western blotting using this antibody. Intracellular canine SIRT1 was also detected in permeabilized 293T cells transfected with a canine SIRT1 expression plasmid by flow cytometry using this antibody. SIRT1 was detected in all leukocyte subsets including lymphocytes, granulocytes and monocytes. The expression level was markedly different among individual dogs. These results indicated that the method applied in this study is useful for evaluating canine SIRT1 levels in PBMCs from dogs.
The life expectancy provides valuable information about population health. The life expectancies were evaluated in 12,039 dogs which were buried or cremated during January 2012 to March 2015. The data of dogs were collected at the eight animal cemeteries in Tokyo. The overall life expectancy of dogs was 13.7 (95% confidence interval (CI): 13.7–13.8) years. The probability of death was high in the first year of life, lowest in the fourth year, and increased exponentially after four years of age like Gompertz curve in semilog graph. The life expectancy of companion dogs in Tokyo has increased 1.67 fold from 8.6 years to 13.7 years over the past three decades. Canine crossbreed life expectancy (15.1 years, 95% CI 14.9–15.3) was significantly greater than pure breed life expectancy (13.6 years, 95%CI 13.5–13.7, P-value <0.001). The life expectancy for male and for female dogs were 13.6 (95% CI: 13.5–13.7) and 13.5 (95% CI: 13.4–13.6) years, respectively, with no significant difference (P=0.097). In terms of the median age of death and life expectancy for major breeds, Shiba had the highest median age of death (15.7 years), life expectancy (15.5 years) and French Bulldog had the lowest median age of death (10.2 years), life expectancy (10.2 years). When considering life expectancy alone, these results suggest that the health of companion dogs in Japan has significantly improved over the past 30 years.
The aim of present study was to identify risk factors among laboratory findings for mortality in calves with diarrhea. A retrospective analysis was conducted utilizing medical records of 221 diarrheic calves (10.4 ± 3.7 days old) with no concurrent severe disorders that were treated with intravenous fluid therapy from the initial examination. Thirty-eight of the diarrheic calves (17.2%) died within 35 days from the initial examination. Multivariate logistic regression analysis indicated that hypoglycemia (OR 3.09; 95% CI 1.22–7.87; P=0.02) and failure of respiratory compensation (OR 2.63; 95% CI 1.05–6.62; P=0.04) were the major risk factors associated with a negative outcome in diarrheic calves. According to the Kaplan-Meyer analysis, diarrheic calves with hypoglycemia and/or failure of respiratory compensation had a significantly shorter survival than calves without these factors.
Localized scleroderma (LS) is a sclerotic skin disorder rarely reported in the veterinary literature. We herein report the first case of a linear LS-like skin lesion in a cat. A 1-year-old castrated male Himalayan cat was presented with a 1-month history of an alopecic, indurated, serpiginous, branched skin lesion on the dorsal cervical to scapular area. The cat had no history of trauma, although a topical spot-on endectocide had been applied near the lesion. Histopathological examination revealed a focal area of hyperplastic dermal collagen with the absence of pilosebaceous units. The cutaneous lesion remained unchanged during a 2-year follow-up period. Clinical and histopathological similarities of this skin lesion with those of the linear form of LS in humans were considered.
The object is to determine the neuroprotective and antioxidative effects of submicron and blended Lycium barbarum (LB) on retinal degeneration as evaluated by ERG, retinal histopathology and assays of antioxidant (total GSH) and peroxidant (MDA) in the retina. A rat model of light-induced retinal degeneration was used to assess the protective effect of different forms of Lycium barbarum (LB) on retinal degeneration. Rats were divided into four experimental groups, normal control, light-induced untreated, submicron LB and blended LB treated. The rats of submicron and blended groups were treated with 250 mg/kg LB orally once daily for 54 days, followed by induction of retinal degeneration. Retinal function was assessed by electroretinography (ERG). Enzyme-linked immunosorbent assay of the retina lysates was measured for the levels of antioxidants, reduced glutathione and glutathione disulfide, and peroxidants, malondialdehyde, in the retina. The ERG results showed a protective effect in LB treated groups with a greater effect observed in submicron LB treated group than the blended LB treated group. There were higher levels of GSH plus GSSG and lower MDA in submicron LB treated group than other groups. In conclusion, LB provided protective and antioxidative effects on the rat retina with light-induced retinal degeneration. Submicron LB protected degenerative retina better than blended LB. LB is effective against oxidative stress in the degenerative retina.
Cardiovascular disease (CVD) has a tremendous impact on the quality of life of humans. While experimental animals are valuable to medical research as models of human diseases, cardiac systems differ widely across various animal species. Thus, we examined a CVD model in cynomolgus monkeys. Laboratory primates are precious resources, making it imperative that symptoms of diseases and disorders are detected as early as possible. Thus, in this study we comprehensively examined important indicators of CVD in cynomolgus monkeys, including arterial blood gas, complete blood count (CBC), biochemistry and cardiac hormones. The control group included 20 healthy macaques showing non-abnormal findings in screening tests, whereas the CVD group included 20 macaques with valvular disease and cardiomyopathy. An increase of red blood cell distribution width was observed in the CBC, indicating chronic inflammation related to CVD. An increase of HCO3 was attributed to the correction of acidosis. Furthermore, development of the CVD model was supported by significant increases in natriuretic peptides. It is suggested that these results indicated a correlation between human CVD and the model in monkeys. Moreover, blood tests including arterial blood gas are non-invasive and can be performed more easily than other technical tests. CVD affected animals easily change their condition by anesthesia and surgical invasion. Pay attention to arterial blood gas and proper respond to their condition are important for research. This data may facilitate human research and aid in the management and veterinary care of nonhuman primates.
Five calves that had shown neurological symptoms within 9 days after birth were histopathologically diagnosed as encephalomalacia. Two calves showed bilateral laminar cerebrocortical necrosis and neuronal necrosis in the corpus striatum and hippocampus. Since the distributional pattern of the lesions was consistent with that of global ischemia in other species, the lesions were probably hypoxic/ischemic encephalopathy consistent with the history of dystocia and perinatal asphyxia. One calf also showed bilateral laminar cerebrocortical necrosis. However, the lesions were chronic ones, because the calf had survived for long time and necropsied at postnatal day 118. Additionally, the lesions did not involve the corpus striatum and hippocampus. The other two calves showed multifocal necrosis with vascular lesions characterized by fibrin thrombi, perivascular edema and perivascular hyaline droplets in the cerebral cortex, corpus striatum, thalamus, brain stem and cerebellum. Considering the age of onsets and histopathological appearance, it was possible that latter three calves were also hypoxic/ischemic encephalopathy, however, exact cause of them was not revealed. In all calves, degenerated/necrotic neurons showed positive reactions for Fluoro-Jade C and degenerated axons showed immunoreactivity for Alzheimer precursor protein A4. Therefore, these markers were applicable to examination of brain injury in neonatal calves.
A 12-year-old intact male Welsh Corgi was presented with enlargement of the right scrotum. Both testicles were surgically removed and histopathologically examined. On gross examination, white nodules were found in the epididymis and ductus deferens. Histopathologically, the nodules developed continuously from the tunica vaginalis testis of the right scrotum and consisted of spindle-shaped neoplastic cells that invaded the surrounding tissue. Immunohistochemically, the neoplastic cells were diffusely positive for vimentin, cytokeratin and Wilms tumor-1 (WT-1). Based on these findings, the tumor was diagnosed as sarcomatoid mesothelioma. The dog presented with respiratory distress 122 days after surgery and clinical examination found multiple metastatic lesions in the lung, abdominal lymph nodes and peritoneum. The dog died 144 days after surgery due to disease progression.
An underweight 10-month-old Holstein heifer presented with anorexia and ananastasia and was euthanized. Postmortem examination revealed extensive ulceration in the esophagus, tongue, and omasum. Histopathological examination revealed severe necrotic esophagitis, glossitis, and omasitis. Many Gram-negative bacilli were detected throughout the necrotic area in the digestive tract; these were identified as Pseudomonas aeruginosa on the basis of isolation tests, molecular examinations, and immunohistochemistry. Gram-negative long filamentous organisms in the superficial layers of the necrotic lesions reacted positively with antibodies against Fusobacterium necrophorum subsp. necrophorum. Thus, the necrotic lesions were confirmed to be associated with P. aeruginosa and Fusobacterium spp. This is the first detection of P. aeruginosa in bovine esophageal and glossal ulcers associated with Fusobacterium spp.
The objective of this study was to evaluate the influence of the dog’s long-term separation from its owner in the novel environment on the occurrence of psychogenic and oxidative stress. Group I dogs (n=9) were brought to the veterinary clinic and stayed in a kennel room for 12 hr before the surgery, and group II dogs (n=9) −for 10 min before the surgery. Physiological parameters (heart rate (HR) (beats/min) and respiratory rate (fR) (breaths/min)) were measured and blood sampling was done 12 hr before the surgery (T0) for group I dogs and 10 min before the surgery (T1) for both groups dogs. Oxidative stress index (OSI) was determined using spectrophotometer and Rel Assay Diagnostics kits by measuring TAS ant TOS in blood plasma. The cortisol level was measured using AIA-360 Automated Immunoassay Analyzer and ST AIA-pack Cortisol assays. Group I dogs’ HR and fR were elevated at T0 and T1, and group II dogs’−at T1 compared to physiological range. OSI and cortisol levels in group I dogs was higher at T1 compared to T0 (P<0.05). There was no significant difference in cortisol level between group I at T0 and group II at T1 (P>0.05). It might be concluded that dogs’ longer stay in the novel environment without the owner induced significant changes in OSI and cortisol level, which could lead to slow wound healing and the occurrence of systemic diseases.
Two inbred strains of Sprague-Dawley rats, known as the Hatano high- and low-avoidance animals (HAA and LAA respectively), have been selectively bred for high versus low rates of avoidance responses in a shuttle-box avoidance task. To investigate differences in the sexual behavior of Hatano rats, male HAA, LAA and SD rats were tested from 12 to 15 weeks of age. LAA rats exhibited more rapid and frequent sexual behavior than HAA or SD rats, and such differences increased with repeated sexual experience. Plasma testosterone levels tended to be lower in LAA rats than in HAA or SD rats, suggesting that active sexual behavior in LAA rats is not related to these levels. Strain differences in mating behavior between HAA and LAA rats may be caused by emotional responses to novelty.
Digital Dermatitis is a localized infectious dermatitis caused by Treponema-like spirochetes. Antibiotics, such as lincomycin, are currently used for treatment, but their use imposes a withdrawal period. This study investigated the therapeutic effect of topical application of the natural component allyl isothiocyanate, in combination with maintenance hoof trimming, on bovine Digital Dermatitis. Study cows were divided into two groups, the Trimming Group and Non-Trimming Group. The day when allyl isothiocyanate was applied, along with hoof trimming, was set as Day 0. Lesion scores, pain, and the presence of Treponema-like spirochetes on the surface of hooves and in biopsy samples of the tissues were evaluated until Day 6. Both groups showed improvement of lesion scores and improved elimination of Treponema-like spirochetes from within the tissues. The presence of Treponema-like spirochetes on the surface of lesions was significantly higher in the Non-Trimming Group by Day 6. These results suggest that allyl isothiocyanate has therapeutic effects on Digital Dermatitis, when combined with hoof trimming, and may prevent a relapse of dermatitis and a re-infection of Treponema-like spirochetes.
Remifentanil is an ultra-short-acting µ-opioid receptor agonist. The purpose of this study was to determine the relationship of the minimum alveolar concentration (MAC) of sevoflurane and other MAC derivatives, including the MAC for blocking adrenergic responses (MAC-BAR) and the MAC at which tracheal extubation is occurred (MAC-extubation), with or without remifentanil infusion. Six healthy adult beagle dogs were randomly anesthetized three times for determining the MAC-BAR (SEVMAC-BAR), MAC (SEVMAC), and MAC- extubation (SEVMAC-extubation) of sevoflurane under infusion of saline and remifentanil at rates of 0.15, 0.30, 0.60, 1.20, and 2.40 µg/kg/min. The ratio of the SEVMAC-BAR and SEVMAC and that of the SEVMAC-extubation and SEVMAC were not significantly different at baseline and during treatment. The MAC-BAR95 and MAC95 decreased in a dose-dependent manner until reaching 1.20 µg/kg/min, and the MAC-extubation5 decreased in a dose-dependent manner until reaching 0.60 µg/kg/min. The percentage reduction of SEVMAC-BAR, SEVMAC, and SEVMAC-extubation increased in a dose-dependent manner during remifentanil infusion. The heart rate significantly decreased in the MAC-BAR and MAC groups, and the systolic and mean arterial pressures increased after remifentanil infusion compared with the baseline values. Remifentanil infusion caused reduction of the SEVMAC-BAR, SEVMAC, and SEVMAC-extubation in a dose-dependent manner, and ceiling effects were observed in the dogs. Higher doses of remifentanil and sevoflurane were necessary for blocking the sympathetic response to the supramaximal stimulus to prevent movement and extubation in dogs.
A 4-year-old, 5.9-kg female Japanese Spitz presented with syncope and exercise intolerance. Echocardiography revealed an ostium primum atrial septal defect (ASD), a cleft mitral valve, mitral valve regurgitation (MR), and tricuspid regurgitation (TR) (velocity: 3.6 m/sec, pressure gradient: 52 mmHg), leading to a diagnosis of partial atrioventricular septal defect (AVSD) with moderate pulmonary hypertension (PH). Open-heart surgery using cardiopulmonary bypass was performed through right atriotomy. The cleft of the mitral valve was sutured with polypropylene and the AVSD was closed using an autologous pericardial patch fixed with glutaraldehyde. No postoperative pulmonary hypertensive crisis occurred. Shunting flow through the ASD, TR and PH had completely disappeared 2 months postoperatively; however, moderate MR persisted. The dog is still alive 5 years postoperatively without clinical signs.
The treatment of cystic ovarian disease (COD) in dairy cows is still controversial, and some researchers recommend using gonadotropin-releasing hormone (GnRH) regardless of the type of cysts. The aim of this study was to comparatively evaluate the reproductive performance of cows diagnosed with follicular or luteal cystic structures, after treatment with either buserelin acetate (GnRH agonist) or dinoprost (prostaglandin F2-alpha or PGF2α). The diagnosis was established by ultrasonographic examinations performed twice a month starting 40–45 days after calving, until the cows were diagnosed pregnant after artificial insemination. Both types of cysts were treated either with 21 µg buserelin acetate or 25 mg dinoprost, resulting in four subject groups. After treatment, the estrus rate in cows with follicular cysts treated with dinoprost (55.1%) was significantly lower (P<0.05) than in cows treated with buserelin acetate (77.5%) or in cows diagnosed with luteal cysts (77.2% for dinoprost and 72.4% for buserelin acetate). The conception rate was higher (P<0.05) in cows with follicular cysts treated with buserelin acetate (67.7%) than in those treated with dinoprost (60%) or in those with luteal cysts (56.9% for dinoprost and 47.5% for buserelin acetate). These results show that it is preferable to differentiate between the two types of ovarian cysts before treatment. Luteal cysts can be treated with either PGF2α or GnRH analogues, with better results when PGF2α is used. Contrastingly, follicular cysts are better treated with GnRH analogues. When differentiation is not possible, GnRH analogues are recommended over PGF2α.
Feline coronavirus (FCoV) is classified into two biotypes based on its pathogenicity in cats: a feline enteric coronavirus of low pathogenicity and a highly virulent feline infectious peritonitis virus. It has been suspected that FCoV alters its biotype via mutations in the viral genome. The S and 3c genes of FCoV have been considered the candidates for viral pathogenicity conversion. In the present study, FCoVs were analyzed for the frequency and location of mutations in the S and 3c genes from faecal samples of cats in an animal shelter and the faeces, effusions, and tissues of cats that were referred to veterinary hospitals. Our results indicated that approximately 95% FCoVs in faeces did not carry mutations in the two genes. However, 80% FCoVs in effusion samples exhibited mutations in the S and 3c genes with remainder displaying a mutation in the S or 3c gene. It was also suggested that mutational analysis of the 3c gene could be useful for studying the horizontal transmission of FCoVs in multi-cat environments.
A long-term animal experiment involving inoculation with bovine coronavirus (BCoV) was conducted to verify its persistent infection in cattle. Three colostrum-deprived Holstein calves were housed separately in individual rooms of a high-containment facility and inoculated with the BCoV strain Kumamoto/1/07. Until the end of the experiment (1,085, 700 and 280 days, respectively), viral RNAs were detected sporadically by RT-PCR and nested PCR from plasma, nasal discharge, and feces. Seroconversion and titer changes were validated by hemagglutination inhibition tests and neutralization tests. Among the samples, nasal discharge showed a higher viral positivity than feces, which seemed to be associated with positive detection in the plasma. These data demonstrate the existence of persistent infection of BCoV in the respiratory tissues of cattle.
A simple, non-alcoholic extraction method for measuring estrogen and progesterone metabolites in excreta using enzyme immunoassays (EIAs) was developed in Japanese macaques. The obtained detection limits of EIAs using estrone conjugates (E1C), pregnanediol glucuronide (PdG), and estriol glucuronide (E3G) polyclonal antibodies with cross-reactivity to urinary and fecal steroid metabolites were 6.6 pg/ml, 2.1 ng/ml and 0.35 ng/ml, respectively. These assays allowed the determination of E1C, PdG, and E3G from the excreta with good reproducibility and accuracy. Thereafter, urine and fecal samples of two menstrual cycles and six pregnancies from eight female Japanese macaques were assayed. A typical increase in urinary and fecal E1C in follicular phase and PdG in luteal phase were shown during non-conceptive menstrual cycles. Urinary E3G levels also showed a preovulatory increase; however, fecal E3G levels were very low throughout the non-conceptive menstrual cycles. Levels of E1C and PdG in the urine and feces of pregnant females were gradually increased until parturition, while fecal E3G levels were low and reached detectable levels after the mid-pregnancy period. Although the extraction rate of estrogen and progestogen metabolites by our method was lower compared to those of the previous extraction method using an alcohol-containing buffer, our method was simple, and the correlation coefficients for the relationship between two methods were found to be statistically significant. The results presented here are of great practical value for a non-invasive method of monitoring ovarian function and pregnancy in Japanese macaques.
Flying foxes are important in the maintenance of forests and diversity. However, knowledge of their behavioral ecology, especially of movement and foraging patterns, which are essential for conservation and management of their populations, are not well known. Therefore, movement behavior of two individuals of Pteropus vampyrus were examined using an Argos telemetry system, and foraging pattern of Pteropus spp. was directly observed, at West Java province, Indonesia in October 2017. The maximum distance between the location at which bats were released and their furthest roost, recorded via satellite telemetry, was approximately 100 km. This reflects the long-distance flight ability of P. vampyrus. Daytime roosting sites and nighttime foraging places consisted of several types of habitats, such as intact forests, agricultural lands, and residential areas. This evidence indicated that there was habitat overlap between humans and bats in West Java province. According to direct observation of the behaviors of flying foxes at two locations within residential areas, various activities such as wing spreading, excretion, fighting, aggressive calls, movement, hanging relax, and hanging alert were found. The number of bat-visits to the trees varied among night hours, and had a positive correlation with the number of fruit dropping. The data obtained in this study have improved our understanding of nighttime behavior and habitat utilization of P. vampyrus, that can be used to support landscape management, species conservation, and disease prevention in regions of Southeast Asia.