Inactivation of koi‐herpesvirus in water using bacteria isolated from carp intestines and carp habitats

Since its first outbreak in Japan in 2003, koi‐herpesvirus (KHV) remains a challenge to the carp Cyprinus carpio L. breeding industry. In this study, inactivation of KHV in water from carp habitats (carp habitat water) was investigated with the aim of developing a model for rapidly inactivating the pathogen in aquaculture effluent. Experiments with live fish showed that, in carp habitat water, KHV lost its infectivity within 3 days. Indications were that inactivation of KHV was caused by the antagonistic activity of bacteria (anti‐KHV bacteria) in the water from carp habitats. Carp habitat water and the intestinal contents of carp were therefore screened for anti‐KHV bacteria. Of 581 bacterial isolates, 23 showed anti‐KHV activity. An effluent treatment model for the disinfection of KHV in aquaculture effluent water using anti‐KHV bacteria was developed and evaluated. The model showed a decrease in cumulative mortality and in the number of KHV genome copies in kidney tissue of fish injected with treated effluent compared with a positive control. It is thought that anti‐KHV bacteria isolated from the intestinal contents of carp and from carp habitat water can be used to control KHV outbreaks.     

Histopathological comparison of pulmonary artery lesions between raccoon dogs (Nyctereutes procyonoides) and domestic dogs experimentally infected with Dirofilaria immitis

Five raccoon dogs (Nyctereutes procyonoides) and two domestic dogs (Canis familiaris) were subcutaneously infected with 100 infective larvae (L3) of Dirofilaria immitis. Two and five worms, respectively, were collected from two of three raccoon dogs. Villous endarteritis was found in the raccoon dog with five worms and two dogs at 116 days after infection. The number of recovered worms in the raccoon dogs was significantly smaller than that of the domestic dogs having 22 and 29 worms, while histopathological features and the severity of the lesions in the raccoon dogs were similar to those in the domestic dogs. The vascular lesions in two chronically-infected raccoon dogs turned into much severe at 565 and 590 days after inoculation.     

Ghrelin differentially modulates the GH secretory response to GHRH between the fed and fasted states in sheep

The effect of energy balance on the growth hormone (GH) secretory responsiveness to growth hormone-releasing hormone (GHRH) has not been determined in ruminant animals. Therefore, we examined the effects of intravenous injections of 0, 3.3, and 6.6 μg ghrelin/kg body weight (BW), with and without GHRH at 0.25 μg/kg BW, on GH secretory responsiveness in both the fed and fasted sheep. The injections were carried out at 48 h (Fasting state) and 3 h (Satiety state) after feeding. Blood samples were taken every 10 minutes, from 30 minutes before to 120 minutes after the injection. Low (3.3 μg/kg BW) and high (6.6 μg/kg BW) doses of ghrelin stimulated GH secretion significantly (P < .05) greater in the Satiety state than in the Fasting state. Growth hormone-releasing hormone plus both doses of ghrelin stimulated GH secretion significantly (P < .05) greater in the Satiety state than in the Fasting state. Ghrelin and GHRH exerted a synergistic effect in the Satiety state, but not in the Fasting state. Plasma ghrelin levels were maintained significantly (P < .05) greater in the Fasting state than in the Satiety state except the temporal increases after ghrelin administration. Plasma free fatty acid (FFA) concentrations were significantly (P < .01) greater in the Fasting state than in the Satiety state. In conclusion, the present study has demonstrated for the first time that ghrelin differentially modulates GH secretory response to GHRH according to feeding states in ruminant animals.     

Optimization of Ca2+ concentrations in fusion and activation media for production of cloned embryos from miniature pig somatic cells

The effects of Ca(2+) concentration in activation medium and cytochalasin B treatment after activation on the parthenogenetic development of pig oocytes were examined. In addition, cloned embryos derived from miniature pig somatic cells were activated under optimal conditions and the effects of Ca(2+) in fusion medium on the development of embryos after activation was examined. When oocytes were activated in 0.1 mM Ca(2+) and then treated with cytochalasin B, the blastocyst formation rate (28.6%) was significantly higher than those activated in 0-0.05 or 1.0 mM (11.0-18.3%). Treatment with cytochalasin B decreased the second polar body extrusion rate of activated oocytes. The presence or absence of Ca(2+) in fusion medium did not affect the fusion rate of miniature pig somatic cells with recipient oocytes. A few cloned embryos developed to the blastocyst stage (2.7-9.0%) without an additional activation treatment. On the other hand, significantly more embryos developed to the blastocyst stage after activation treatment when they were fused in the absence (28.9%) of Ca(2+) rather than the presence (16.5%) of it. These results show that the highest blastocyst formation rate for miniature pig cloned embryos is obtained when donor cells and recipient oocytes are fused in the absence of Ca(2+) and then activated in 0.1 mM Ca(2+) and treated with cytochalasin B.     

Some Characteristics of Primary and Secondary Oestrous Signs in High-producing Dairy Cows

Shortened and weakened oestrous signs in dairy cows may cause a failure of oestrus detection and artificial insemination timing error leading to poor reproductive performance. The aims of this study were to investigate the duration of standing oestrus in high-producing dairy cows under a free stall system, to determine the duration of expression of secondary oestrous signs before and after standing oestrus (Expt 1) and to compare the duration and intensity of oestrus between cows and heifers (Expt 2). Cattle were checked for primary and secondary oestrous signs at an interval of 4 h. Heat detection aids were also used. In Expt 1, of 56 cows which were detected in oestrus, 36 cows (64.3%) showed standing oestrus and other 20 cows (36.6%) showed secondary oestrous signs only. Duration of the standing oestrus was 6.6 +/- 6.3 h on average (+/-SD), ranging between 2 and 32 h. The cows in standing oestrus showed secondary oestrous signs during a period from 9.6 +/- 8.1 h before onset of standing to 18.4 +/- 18.8 h after the end of standing oestrus. In the cows that did not show standing oestrus, expression of secondary oestrous signs were observed for 25.7 +/- 20.5 h, which was 7.5 h shorter than the average duration of oestrus in cows showing standing oestrus. In Expt 2, nine (82%) of the 11 lactating cows in oestrus showed standing, while all the 10 heifers exhibited standing oestrus. Average duration of standing oestrus was 6.4 +/- 4.3 h in cows and 6.2 +/- 3.9 h in heifers, respectively. It may be concluded that the duration of standing oestrus is substantially shortened in lactating dairy cows, and more than one-third of cows did not show standing oestrus. In cows showing standing oestrus, duration of expression of secondary oestrous signs before and after standing is not shortened. Duration of standing oestrus in heifers was as short as that in cows.     

Response of plasma cortisol and progesterone after ACTH challenge in ovariectomized lactating dairy cows

Shortened and weakened estrous expressions could be one of the causes of poor heat detection rate. Non-specific acute stresses are assumed to depress expression of estrus by an increase of plasma progesterone which may originate from the adrenal cortex. The objective of the present study was to examine whether the adrenal cortex can secrete significant amounts of progesterone in response to exogenous adrenocorticotropic hormone (ACTH) in lactating cows. Four cows had estrus synchronized and were ovariectomized in the luteal phase. The cows were given 25 IU ACTH through an indwelling catheter 5 h after catheterization. Blood samples were collected at an interval of 30 min. In 3 of the 4 cows, plasma progesterone concentrations increased significantly 0.5-1.5 h after the first ACTH challenge with a mean peak value of 4.2 +/- 0.4 (S.D.) ng/ml. A similar response was also observed after the second ACTH challenge. Peak plasma progesterone concentrations in the 3 cows after first ACTH challenge were comparable with the progesterone values in the luteal phase of each cow. The results suggest that lactating cows have the capability to secrete a significant amount of progesterone from the adrenal cortex.     

Feather-pulp lesions in chickens with naturally occurring Marek's disease lymphomas

Feather pulps of 15-to-35-week-old chickens with Marek's disease (MD) lymphomas were examined histopathologically. Of the 64 chickens, 59 (92.2%) had lymphoproliferative (T-type) lesions in the feather pulp. The T-type feather-pulp lesion (FPL) occurred in all regions, but more frequently in the upper column of the pulp. Severe lesions were distributed throughout the feather pulp. Some of the T-type FPLs regressed to inflammatory lesions consisting of necrosis or loss of constituent cells, edema, and infiltration by small lymphocytes, heterophils, and plasma cells. The regressive T-type FPL was usually diffuse throughout the tumorous lesions, but proliferative foci were often seen concomitantly with regressive lesions. The grade and histologic picture of T-type FPLs in chickens correlated well with those of the visceral lymphomas. These findings suggest that the severity of MD lymphoma is predictable without autopsy by examining feather-pulp samples.     

Isolation of Staphylococcus aureus from raw fish in relation to culture methods

Five hundred and fifty fish samples from various stages in the course of distribution in Hyogo Prefecture (209 retailed in super markets, 173 obtained from fishery cooperatives at a harbor, 91 caught by trawling and 77 caught by rod fishing) were examined for contamination with Staphylococcus aureus (S. aureus). S. aureus was detected in 41 (19.6%) of the retail fish samples and 46 (26.6%) of the samples from the fishery cooperatives. No S. aureus was isolated from the live fish (91 trawled and 77 fished by rod). With regard to the retail fish, the contamination rate of processed fish (26.0%) was significantly higher than that of unprocessed fish (14.2%). For 88 samples, the efficacy of the selective medium was compared using Baird-Parker agar and mannitol salt agar supplemented with egg yolk (MSEY agar) by the direct plate and enrichment culture methods. Using the direct culture method, the S. aureus positive rate with the Baird-Parker agar (30.7%) was significantly higher (P<0.01) than that with the MSEY agar (6.8%). The enrichment culture method remarkably raised the S. aureus detection rate. Seventy-eight (85.7%) of 91 isolates belonged to the human ecovar. Sixty-two (68.1%) of the 91 isolates had some enterotoxin genes, including 44 (48.4%) with the sea gene. These data showed that the fish were contaminated with S. aureus after landing and that Baird-Parker agar had an advantage in detecting S. aureus with a direct plate culture.     

The banding patterns of normal canine chromosomes

The chromosomal banding patterns of canine normal chromosome were analyzed by G-, Q-, C- and N-banding techniques. The chromosomes of canine normal lymphocytes consisted of 38 acrocentric autosome pairs and X-, Y-metacentric sex chromosomes. Their G- and Q-banding patterns corresponded to the those reported by Manolache et al. in 1976. C-bands of chromosomes were found on the centromeric regions of the 7th, 26th, 34th, 35th, 37th and 38th pairs of chromosomes. In addition, N-bands whose location had not been confirmed until now were recognized on the telomitic regions of the 7th, 9th and 22nd pairs of chromosomes.