Chronic otitis externa with heat shock protein 70-positive intranuclear inclusion bodies in the ceruminous gland epithelium of a Chihuahua dog

A Chihuahua dog showed persistent itching in the right ear canal. Anti-inflammatory medicines and prednisolone were ineffective and total ear canal ablation was performed. Histological diagnosis was chronic otitis externa. Eosinophilic intranuclear inclusion bodies (Cowdry type A and full-type) were occasionally observed in the ceruminous gland epithelium. The inclusion bodies were negative for nucleic acid and ultrastructurally composed of fibrous structures (approximately 10 nm in width). Viral infection was initially suspected, but polymelase chain reaction tests did not detect the expected viral genes. Immunohistochemistry revealed that the inclusion bodies were positive for heat shock protein 70 (HSP70), suggesting that these bodies could be protein aggregates including HSP70. The etiology of this lesion has not been elucidated, but chronic inflammation may influence the cytoplasm-to-nuclear transportation of HSP70. To the best of our knowledge, this is the first report of canine chronic otitis externa with HSP70-positive intranuclear inclusion bodies.     

Phylogenetic Analysis of Field Isolates of Feline Calcivirus (FCV) in Japan by Sequencing Part of Its Capsid Gene

The molecular epidemiology of the infectious disease caused by feline calcivirus (FCV) in Japan was investigated by analysing the phylogenetic relationship among 21 Japanese field isolates, including the F4 strain, and 30 global isolates. Parts of the capsid gene (B–F) of the isolates were amplified by RT-PCR, and the amino acid sequences were compared with those from the global isolates. Thirty-seven and 14 out of a total of 51 isolates were clustered into two distinct genogroups, I and II respectively, by UPGMA and NJ analysis. Seven of the 21 Japanese isolates (33%) fell into group I together with 30 global isolates, while the other 14 Japanese isolates (67%) belonged to group II. The bootstrap repetition analysis of groups I and II formed by the NJ method gave a value of 99.0%. The 14 latter Japanese isolates were clearly separated from the isolates in group I, and they were different from any previously known FCV, forming a new genogroup, which implies that this lineage has been confined to Japan. Comparing the amino acid sequences shared by groups I and II, the amino acid at position 377 in B region was asparagine (Asn or Asp (NH₂)) in group I, while it was lysine (Lys) in all the strains in group II. Similarly, the amino acid at position 539 in the F region was alanine (Ala) or proline (Pro) in group I, while it was valine (Val) in group II; glycine (Gly) at position 557 in group I was serine (Ser) in Group II; and phenylalanine (Phe) or leucine (Leu) at position 566 in genogroup I was tyrosine (Tyr) in group II.     

Effects of photoperiod on salsolinol‐induced prolactin secretion in goats

The aim of the present study was to clarify the relation between salsolinol (SAL)‐induced prolactin (PRL) release and photoperiod in goats. A single intravenous (i.v.) injection of SAL was given to adult female goats under short (8 h light, 16 h dark) or long (16 h light, 8 h dark) photoperiod conditions at two different ambient temperatures (20°C or 5°C), and the PRL‐releasing response to SAL was compared to that of thyrotropin‐releasing hormone (TRH) or a dopamine (DA) receptor antagonist, sulpiride. SAL, as well as TRH or sulpiride, stimulated the release of PRL promptly after each injection in both 8‐ and 16‐h daily photoperiods at 20°C (P < 0.05). The area under the response curve (AUC) of PRL for the 60‐min period after injections of saline (controls), SAL, TRH and sulpiride in the 16‐h daily photoperiod group was greater than each corresponding value in the 8‐h daily photoperiod group (P < 0.05). There were no significant differences in the AUC of PRL among the values produced after the injection of SAL, TRH and sulpiride in 16‐h daily photoperiod group; however, the values produced after the injection of TRH were smallest among the three in the 8‐h daily photoperiod group (P < 0.05). The PRL‐releasing responses to SAL, TRH and sulpiride under a short and long photoperiod condition at 5°C resembled those at 20°C. These results show that a long photoperiod highly enhances the PRL‐releasing response to SAL as well as TRH or sulpiride in either medium or low ambient temperature in goats.     

Morphological observation of canine natural killer cells mediated cytotoxicity.

The cytotoxic effects of canine NK cells on CL-1 target cells were examined by scanning electron microscopy (SEM). NK cell mediated cytotoxicity on CL-1 target cells was detected by 51Cr release assay. SEM showed that a canine NK cell extended projections to the CL-1 target cell. Furthermore, the surface of CL-1 target cells changed a mesh-like structure. Therefore, the cytotoxic effects of canine NK cells on CL-1 target cells were morphologically demonstrated.     

Degradation of the urban ecosystem function due to soil sealing: involvement in the heat island phenomenon and hydrologic cycle in the Tokyo metropolitan area

ABSTRACT

We reviewed the role of the soil function of urban green space in mitigating the heat island phenomenon and urban flood damage, which are important issues in the modern urban environment. Urban sprawl has progressed remarkably in the southern part of the Kanto District of Japan, especially since the 1960s. The grassland/bare land area ratio in the center of Tokyo was more than 70% in the 1930s but less than 40% in 1990. On the other hand, the paving area ratio was ~2% in the 1930s but more than 10 times that amount in 1990. Thus, cities, such as Tokyo, in the southern part of the Kanto District have been significantly sealed soil throughout time, and the heat island phenomenon has intensified. Urban green space helps to mitigate the heat island phenomenon based on the water retention and heat dissipation function of plants and unsealed soil. A cool island effect was reported during summer days in urban green spaces in Tokyo. The mitigation of the heat island effect seems to be large, even when conditions, such as the park area, land use, afforestation area ratio, and soil type, differ. Soil sealing and compaction affect urban flooding by hindering the penetration of rainwater and increasing surface runoff. According to a survey in the hilly basin (1 km²) of the western part of Tokyo, the proportion of farm- and forestland decreased due to development. The urban area increased from ~10% in the 1970s to ~60% in the 1980s. As a result, the flood arrival time shortened. Green spaces have a high rainwater permeability. The forest soil structure and presence of O horizons further increase the permeability. By promoting permeability to the underground, it is possible to reduce and delay the water runoff on the land surface. To develop urban green space as green infrastructure, it is necessary to accumulate more information on the current situation and agenda for the future of urban green space.     

Cryopreservation of canine spermatozoa using a skim milk-based extender and a short equilibration time

Although useful spermatozoa cryopreservation techniques have been established, long-term equilibration seems to be required before freezing the spermatozoa of many species, including dogs. The fertility of cryopreserved dog spermatozoa from five males for a reduced equilibration period (0, 30, 60, 120 and 180 min) in a skim milk (SM)-based extender containing raffinose was evaluated in the present study. When the sperm was diluted with the extender at room temperature (RT) and cryopreserved without equilibration, the proportion of total motile spermatozoa (TMS) after thawing was lower (27%) than when the sperm was equilibrated for 30 min (33%), 60 min (32%), 120 min (44%; p < .05) or 180 min (29%). The proportion of TMS increased as the equilibration time increased and peaked at 120 min. Acrosome integrity was significantly lower in the cryopreserved spermatozoa that had not undergone the initial equilibration than in the equilibrated spermatozoa (p < .05). The normal rate of acrosomes increased with the extension of the first equilibration and peaked at 120 min. When frozen–thawed spermatozoa that had been diluted at RT and subjected to an initial equilibration lasting 60 or 180 min were transcervically inseminated into recipients, there were no differences in the delivery rate, litter size or breeding efficiency. In the cryopreservation of canine spermatozoa using a SM-based extender, even if the initial equilibration time was shortened to 60 min, the results were comparable to those obtained when the conventional method (with an initial equilibration time of 180 min) was used.     

Length of Incubation for the Estimation of the Most Probable Number of Nitrifying Bacteria in Soil

Abstract

The Zn content in 8 soil chemical fractions was determined for 21 greenhouse soils and for 8 reference (open field) soils from Kochi Prefecture, Japan to investigate the forms of spontaneously accumulated Zn in the greenhouse soils associated with heavy application of fertilizers and manures. Sequential extraction method was applied to every soil and each Zn fraction was designated as exchangeable (Ex-Zn), Pb-displaceable (Pb-Zn), acid soluble (Aci-Zn), Mn oxide-occluded (MnO-Zn), organically bound (OM-Zn), amorphous Fe oxide-occluded (AFeO-Zn), crystalline Fe oxide-occluded (CFeO-Zn), and residual (Res-Zn) fractions. The Zn content of the greenhouse soils was significantly higher than that of the reference soils in every fraction, except for the CFeO-Zn, and Res-Zn fractions. The Pb-Zn, Aci-Zn, and MnO-Zn fractions showed a difference of more than 60% in the total Zn content between the two soil groups. The amounts of Zn extracted in the Pb-Zn, Aci-Zn, MnO-Zn, AFeO-Zn, and CFeO-Zn fractions of the greenhouse soils increased con-comitantly with the accumulation of applied macro-nutrients. These results indicated that the accumulation of Zn in greenhouse soils caused by intensive fertilization had proceeded through specific adsorption onto or occlusion by the oxides and hydroxides of Fe and Mn in soils.     

Prevalence of canine coronavirus (CCoV) in dog in Japan: detection of CCoV RNA and retrospective serological analysis

We collected rectal swabs from dogs in Japan during 2011 to 2014, and canine coronavirus (CCoV) nucleocapsid gene was detected by RT-PCR. The relationship between CCoV infection and the manifestation of diarrhea symptoms was investigated, and a correlation was noted (df=1, χ²=8.90, P<0.005). The types of CCoV detected in samples from CCoV-infected dogs were CCoV-I in 88.9% and CCoV-II in 7.4%, respectively. We retrospectively investigated the seroprevalence of CCoV-I in dogs in Japan during 1998 to 2006. The sera were tested with a neutralizing antibody test. In the absence of CCoV-I laboratory strain, we used feline coronavirus (FCoV)-I that shares high sequence homology in the S protein with CCoV-I. 77.7% of the sera were positive for neutralizing anti-FCoV-I antibodies.     

Artificial insemination with canine spermatozoa frozen in a skim milk/glucose-based extender

Due to the recent outbreak of avian influenza, transportation of frozen canine semen with egg yolk has been sharply restricted. Thus, there is urgent need to develop a novel egg yolk-free extender for freezing canine spermatozoa. In the present study, the effect of using skim milk/glucose (SG)-based extender without egg yolk on the motility and fertilizing capacity of canine spermatozoa frozen-thawed in the presence of glycerol was examined. There was a tendency for the proportion of motile spermatozoa exposed to SG-based extender for 3 h to be higher than that exposed for 1 h, but the difference was not significant. The motility and other viability parameters of canine spermatozoa after thawing were similar to those obtained with an egg yolk-based extender. When spermatozoa frozen with SG-based extender containing glycerol after 3 h exposure were transcervically inseminated into 2 recipient bitches, a total of 6 pups were obtained. These results suggest that a simple extender composed of skim milk, glucose and glycerol is useful for cryopreservation of canine spermatozoa, which may contribute to improved exchange of genetic material and efficient production of companion and working dogs, such as guide dogs for the blind.