The principal center of the accessory olfactory system is the accessory olfactory bulb (AOB). In primates, simians are divided into two groups, New and Old World monkeys, and the AOB is present in only New World monkeys. The common marmoset (Callithrix jacchus) is a species of New World monkey. Although the morphology of the common marmoset AOB has been demonstrated, the distribution patterns of the mitral/tufted and granule cells of the AOB remain unclear. In the present study, therefore, the distribution of the mitral/tufted and granule cells in the common marmoset AOB was examined using two histochemical markers including immuno-staining for protein gene product (PGP) 9.5 and NADPH-diaphorase staining. The vomeronasal nerves, gomeruli and mitral/tufted cells showed PGP 9.5-immunoreactivity. The mitral/tufted cells were arranged in only one or two rows along the margin of the glomerular layer to form the mitral/tufted cell layer (MTL). Since the mitral/tufted cells occurred sparsely in the common marmoset, the MTL was illegible. NADPH-diaphorase reactivity was primarily detected in the rostral and caudal areas of the AOB. In these areas, granule cells showed NADPH-diaphorase reactivity. Since the granule cells were sparse, the common marmoset AOB displayed less-developed granule cell layer. Although the functional significance of the AOB remains to be solved in the common marmoset, small-sized and less-laminated AOB may show that sexual behavior of the common marmoset has lesser dependence on the accessory olfactory system.
The adjuvant effect of chicken interferon-γ (ChIFN-γ) was examined for protecting chickens against intestinal colonization of Salmonella Enteritidis (SE) following oral exposure. Ten 7-week-old chickens per group were immunized with inactivated SE twice with or without co-administration of ChIFN-γ intramuscularly, and all chickens were challenged with SE. Sera collected from immunized groups with or without ChIFN-γ, and from unimmunized group were measured for SE antibody by agglutination test. The levels of antibodies were raised by 1 week post-immunization and did not show any difference between groups with and without ChIFN-γ. No antibodies were detected in unimmunized group before challenge. Fecal samples from each group were cultured at 1, 4, 7, and 13 days post-challenge to determine the incidence of intestinal colonization and the numbers of SE shed into the environment. Co-administration of ChIFN-γ, significantly reduced the incidence of intestinal colonization (P<0.05). At 13 days post-challenge, the bacterial counts of SE in organs were also reduced in ChIFN-γ administered group. These data suggest co-administration of ChIFN-γ with SE antigen enhances protection against SE challenge without acceleration of antibody production.
The in vitro suscetibilities of six commonly used antimicrobial agents against 29 isolates of intestinal spirochetes isolated from dogs in Japan were examined by the agar dilution technique. In addition, the genetic basis of tylosin resistance in in vitro selected resistant mutants of two reference strains and three tylosin-susceptible field isolates obtained by three successive subcultures on blood agar containing 1 μg/ml of tylosin was investigated. Carbadox was the most active (MIC: < 0.00625) of all the antimicrobial agents. Although all the isolates were susceptible to tylosin, some were resistant to erythromycin. Tiamulin, lincomycin and dimetridazole were also very active against the isolates. All the resistant isolates did not harbor any plasmids. In vitro selected tylosin-resistant mutants of previously tylosin-susceptible isolates showed a new mutation in which their adenine at the base position equivalent to 2,062 of 23S rDNA of Escherichia coli has been replaced by cytosine. These findings may both provide guidance towards the proper choice of antimicrobial agents for the treatment of canine intestinal spirochetosis, and add to the understanding of the genetic basis of tylosin resistance.
Rhodococcus equi was isolated from fecal and soil samples from four native Jeju horse farms and six Thoroughbred farms in Jeju, Korea. The isolates were examined for the presence of virulence-associated 15-17-kDa antigens (VapA) by colony blotting, using the monoclonal antibody 10G5, and for the gene encoding VapA by PCR. R. equi was isolated from all 36 soil samples collected from the 10 farms with between 5.0 × 102 and 7.5 × 104 colony-forming units (cfu) per gram of soil, and from 37 of 40 fecal samples with between 5.0 × 101 and 1.1 × 10 5 cfu per gram of feces. Virulent R. equi was isolated from seven farms and appeared in 2.0% of isolates (10 of 508). Of the 10 virulent isolates, four contained a 90-kb type II plasmid, which has been found in isolates from the Kiso native horses of Japan, and the other six contained a new variant, which did not display the EcoRI and EcoT22I digestion patterns of the 10 representative plasmids already reported (85-kb types I, II, III, and IV; 87-kb types I and II; 90-kb types I, II, III, and IV). We designated the new variant as the "90-kb type V" plasmid, because its EcoRI digestion pattern is similar to that of the 90-kb type II plasmid. This is the first report of the prevalence of virulent R. equi in Jeju, Korea. The same virulence plasmid type is found in both Korean and Japanese isolates, providing insight into the origin, ancestry, and dispersal of native horses in Korea and Japan.
Eight splenectomized calves were inoculated with Theileria orientalissergenti (Tos)-infected tick gland homogenate (5 calves) or infected erythrocyte suspension (3 calves). Clinical characteristics were different in calves post-infection. Animals were divided into 3 groups on the basis of susceptibility as high, middle, and low. Increase in mRNA of IFN-γ, IL-2, and TNF-α was observed in peripheral blood mononuclear cells at the peak of infection and was seen to be related with pyrexia and parasitemia. Expression of IL-1, IL-4, and inducible nitric oxide synthase was not observed. Decreased plasma nitrite/nitrate level was observed in the groups. The results of this study indicate that Th1 response is the predominant response in Tos infection, and this response is also related with their clinical characteristics.
Ultrasonic vocalizations (USVs) are observed along with freezing behavior when rats are placed under fearful situation. Coping style with stress is categorized into two forms, namely passive and emotional avoidance and active operant avoidance. In this study, fear-induced USV was compared between two conditioning protocols. Two groups of male Wistar rats were contextually conditioned for 10 days by exposing to the shock box. One group was shock-unavoidable and the other shock-avoidable by operant control of emitting the USVs. They were then proceeded to the extinction phase. As the results, the shock-unavoidable group emitted greater USV in both conditioning and extinction phases. The extinction burst, commonly seen in active learned responses, was not observed in either group. The duration of freezing was also longer in shock-unavoidable conditioned rats. These results suggest that under the shock-unavoidable condition, rats receive more stress and thus emit more USV. It is therefore concluded that fear-induced USV is an emotional and passive response to the stress, rather than being controlled in an operant-way.
Early postnatal experiences have been shown to have a tremendous influence on behavior development. In this study, we focused on weaning time and investigated the effects of early weaning on adulthood aggressive behavior in mice with the resident-intruder test. Early weaning resulted in decreased sideways threat and tail rattle. In addition, the frequency of attack bites was more variable in the early weaned group, although the mean frequency did not differ from the normally weaned group. The results of the present study suggest that aggressive behavior is also affected by early weaning manipulation.
The bottlenose dolphin interleukin (IL)-8 cDNA was molecularly cloned. The dolphin IL-8 has an open reading frame of 303-bp encoding 101 amino acids. The homology of the amino acid sequence with that of other species was: sheep, 89.1%; cattle, 88.1%; pig, 85.1%; dog, 85.1%; horse, 79.2%; human, 74.5%; and macaque, 72.3%. The amino acid sequence suggested that dolphin IL-8 was a CXC chemokine. The recombinant dolphin IL-8 protein was recognized with anti-ovine IL-8 monoclonal antibody.
Erythrocyte 5'-nucleotidase is thought to be involved in the maturation of erythrocytes. In the present study, in vitro incubation of canine erythrocytes demonstrated that significant inhibition of 5'-nucleotidase activity occurred in the presence of serum from dogs infected with Babesia gibsoni, when the enzyme was assayed with cytidine 5'-monophosphate (5'-CMP) and inosine 5'-monophosphate (5'-IMP) as substrates. The multiplication of B. gibsoni in in vitro culture also resulted in a significant decrease in the enzyme activity of erythrocytes in the culture. Furthermore, the infected serum and 5'-CMP retarded the maturation of canine reticulocytes in vitro. These results suggested that nucleotides such as 5'-CMP and 5'-IMP might accumulate in young erythrocytes and/or serum in dogs infected with B. gibsoni as a result of decreased activity of erythrocyte 5'-nucleotidase, resulting in the delayed maturation of reticulocytes.
A 22-month-old male Japanese mongrel cat with a history of dysuria and recurrent generalised tonic-clonic seizure was examined by neuro-opthalmological testing and computed tomography (CT). Vision testing revealed narrowing of the visual field in the right eye, and complete visual deficits in the left eye. Pupillary reactions, and motor and sensory function in the eyelids and the eyes were normal. The cat was diagnosed as hydrocephalus by CT examination, because dilation of the right lateral ventricle, and compression of the right temporal and occipital cortices was shown. The etiology of the hydrocephalus was unclear. Although a unilateral lesion of the upper visual pathway was suspected, a complete homonymous hemianopsia was not shown.
The objectives of this study were to evaluate and compare the effects of intravenously (IV) administered infusion of isotonic solution (ISB) or hypertonic sodium bicarbonate solution (HSB) on acid-base equilibrium and the plasma osmolarity in acidemic calves experimentally induced by 5 M-NH4Cl, IV infusion (1.0 ml/kg, over 1 hr). The ISB and HSB infusion induced progressive and significant increases in their HCO3- and BE levels that persisted throughout the period of fluid administration. The plasma osmolarity in the ISB groups was significantly decreased. The plasma osmolarity in the HSB group was significantly higher than in the calves in the other groups (p<0.05). ISB solution might be safe and effective for treating and reviving conscious calves from experimentally induced metabolic acidosis.
To investigate temporal dynamic changes in the synthesis of chondroitin 6-sulfate (CS6) and chondroitin 4-sulfate (CS4) in vitro, normal articular cartilage of femoral heads was harvested from three dogs. Chondrocytes were isolated and cultured in alginate microspheres for 21 days. On days 7, 14 and 21, DNA content was quantified by fluorometric assay using Hoechst 33258. On days 14 and 21, proteoglycans were extracted, and the amounts of CS6 and CS4 were quantified after chondroitinase ABC digestion using capillary electrophoresis. The DNA content and amounts of CS6 and CS4 increased during the culture period. The amounts of CS6 and CS4 divided by DNA content revealed that the synthesis of CS6 was more up-regulated than CS4.
The rectal temperature obtained using a standard electronic thermometer was compared with ear, back skin, tail skin, and sole skin temperatures obtained using an infrared thermometer in B6C3F1 mice. Using both methods, we investigated baseline temperatures, diurnal and 2-week variations in temperatures, and ethanol-induced hypothermia in these body locations. Ear and back temperatures were shown to be close to and consistent with rectal temperatures in various situations, and measured temperatures at these sites were almost constant, with very similar diurnal variation. Conversely, tail and sole temperatures were lower and much more variable. These results indicate that ear and back skin temperatures obtained using a convenient and non-invasive infrared thermometer are as reliable, and should be safer and less stressful to animal subjects, compared to standard rectal temperature measurements.
Lactoferrin (LF), a member of the transferrin (TF) protein family, is an iron-binding protein that is known to interact with bacteria through a specific receptor. We examined the binding of bovine LF (bLF), bovine TF (bTF), and ovotransferrin (OTF) by Toxoplasma gondii using a fluorescence test and the streptavidin-biotin (SAB) method using biotin-streptavidin, and found that bLF, bTF, and OTF bound to the protein components of T. gondii. Furthermore, we confirmed that bLF, bTF, and OTF bound a 42 kDa soluble protein of T. gondii by far Western blot method. These results demonstrated that bLF binding proteins are present on T. gondii.
Histiocytic proliferative disorders in canine central nervous system (CNS) including granulomatous meningoencephalomyelitis (GME) and malignant histiocytosis were compared pathologically. Lesions of GME mainly existed in the white matter of the cerebrum, brainstem and cerebellum and consisted of characteristic perivascular cuffing, parenchymal granuloma and leptomeningeal infiltrates of mononuclear cells. In malignant histiocytosis, there were two histological patterns, diffuse proliferation of neoplastic histiocytes through the leptomeninges and neoplastic nodule formation in the parenchyma. Neoplastic histiocytes exhibited mild to severe cellular atypia and high ability of invasion into the brain parenchyma. Mitotic and phagocytic figures were also observed. Several histiocytic markers, including lysozyme, α1-antitrypsin and lectin RCA-1, revealed histiocytic origin of both inflammatory and neoplastic cells, however, those were not determinative for the discrimination between GME and malignant histiocytosis. CD3- and PCNA-positive cells existed in the lesions of both diseases. The number of CD3-positive cells in GME tended to be greater than in malignant histiocytosis, while the difference was not statistically significant.
CD38 is an ectoenzyme with ADP-ribosyl cyclase and hydrolase activities, which synthesizes cyclic ADP-ribose from NAD and hydrolyzes cyclic ADP-ribose to ADP-ribose. It has been shown that cyclic ADP-ribose is a potent Ca2+ mobilizing messenger in many cells. To know the physiological role of cyclic ADP-ribose in vascular smooth muscle, we examined the effects of various agonists in the aorta isolated from CD38 knockout (CD38-/-) mouse. Western blot analysis showed that CD38 protein was detected in the aorta isolated from wild-type (CD38+/+) mouse, but not from CD38-/- mouse. In the aortae isolated from both CD38+/+ and CD38-/- mice, KCl, phenylephrine and norepinephrine induced concentration-dependent contraction. KCl produced similar concentration-dependent responses in the aortae from both CD38+/+ and CD38-/- mice. Maximum force of contraction induced by KCl (65 mM) was same in the size. Phenylephrine- and norepinephrine-induced contractions were, however, significantly smaller in the aortae from CD38-/- mice than in those from CD38+/+ mice. 5-Hydroxytryptamine, endothelin-1, caffeine and thapsigargin-induced contractions were not significantly different in these two aortae. These results suggest that CD38 gene disruption inhibits α-adrenoceptor-induced vascular contractions and cyclic ADP-ribose-mediated signal transduction system is committed in these responses.
The objectives of this study were to observe subsequent reproductive performance of re-serviced females by the number of services within a parity, to measure mean days to re-service and culling intervals, to determine lifetime performance in re-serviced gilts, and to investigate re-serviced females across parities on commercial farms. Reproduction records on 539 U.S.A. farms were used to observe re-serviced females by the number of service groups at the herd level. Farrowing rate decreases by approximately 10%, and re-service occurrence increases by approximately 5% for each increase in the number of services increase within a parity group (P<0.05). Only in parity 0 to 2 groups, average pigs born alive at subsequent farrowing in the second or later service groups were greater than in the first service group (P<0.05), but in parity ≥3, the third or later service groups produced fewer pigs born alive than the other service groups (P<0.05). Lifetime performance and re-service events were observed in 39,945 individual females on the 149 selected farms that had complete 5-year records. Means of days to re-service, first-mating-to-culling intervals in gilts and weaning-to-culling intervals in sows were 46.3 days, 95.2 days, and 48.2 days, respectively. Re-serviced gilts had longer NPD (>50 days), a lower parity at culling (>0.5) and fewer lifetime pigs born alive (>2 pigs) than non-return gilts (P<0.05), but no difference in average pigs born alive per parity was found between re-serviced gilt groups and non-return gilts. Of 19,677 re-serviced females, 35.6% had two or more re-services across parities in pig life, 10.6% had 3 or more re-services, and 1.95% had four or more re-services. Accurate estrus detection with a boar and improved mating techniques on re-serviced females are suggested to improve herd productivity.
Two azoospermic dogs with high plasma estradiol-17 β (E2) levels were subcutaneously injected with an aromatase inhibitor (AI), 4-androstene-4-ol-3,17-dione, 2 mg every other day for 4 weeks. Before the AI treatment the plasma E2 levels of the two dogs (21 and 22 pg/ml, respectively) were higher than those of 2 normal dogs (8.1 and 12.3 pg/ml), and they fell to 11-17 pg/ml between 1 and 4 weeks after the start of AI treatment. The plasma testosterone levels after the start of AI treatment had increased to 2.1-3.1 ng/ml. A small number of sperm were detected in the semen of the two dogs between 3 and 6 weeks after the start of AI treatment. These results indicate that the testicular function of infertile dogs with high plasma E2 levels can be temporarily improved by AI therapy.
The action of estrogen on target organs has been actively studied with the discovery of estrogen receptor (ER) β. This study was carried out to examine the expression of ERα and ERβ in the uterus and the vagina of immature Sprague-Dawley rats treated with 17-ethinyl estradiol (EE). Twenty days old rats were subcutaneously treated with EE at the doses of 0 (vehicle control), 0.03, 0.3, 1.0, 3.0, and 10.0 μg/kg/day for three consecutive days. The treatment of EE at the doses of 0.3, 1.0, 3.0 and 10.0 μg/kg/day significantly increased the weights of the uterus and vagina of rats (p<0.01) and retained fluid in the uterus of rats. At the high doses of 3.0 and 10.0 μg/kg/day, the treatment of EE caused an increase in the uterine height, hypertrophy, and a decrease in the expression of ERα and ERβ in the uterine luminal and glandular epithelium. The treatment of EE at the doses of 3.0 and 10.0 μg/kg/day also caused cornification and a decrease in the expression of ERα and ERβ in the vaginal epithelium. These results suggest that the EE treatment decrease the expression of ERα and ERβ in the uterus and vagina of immature rats and that may be associated with the morphological changes such as increase in the uterine height, hypertrophy of the uterine epithelium, and cornification of the vagina.
Adverse effects of tributyltin (TBT) chloride were investigated on the reproductive system in male adult rats as exposed during puberty. Fifty Sprague-Dawley rats at the age of 35 days were assigned to five different groups: negative control receiving vehicle, methyltestosterone (10 mg/kg B.W.), and TBT chloride treatments (5, 10, and 20 mg/kg B.W.). Animals were treated by oral gavage for ten consecutive days and sacrificed at 5 weeks after final treatment. The treatment of TBT chloride at the high dose of 20 mg/kg B.W. significantly decreased homogenization-resistant testicular sperm counts (p<0.05). The TBT chloride treatment at the doses of 10 and 20 mg/kg B.W. also significantly decreased caudal epididymal sperm counts (p<0.01). Some of motion kinematic parameters (motility, mean angular displacement, lateral head displacement, and dance) of sperms retrieved from vasa deference were significantly decreased in rats treated with the TBT chloride at the dose of 20 mg/kg B.W. (p<0.05). These results provide a further evidence that an exposure to TBT chloride during pubertal period in male rats produces spermatogenic disorders characterized by decreasing testicular and epididymal sperm counts and some motion parameters of sperms in the vasa deference.