Persistent effect of broody hens on behaviour of chickens

We reported previously that behavioral development of chicks was promoted remarkably by the presence of a broody hen. Here we report that these effects at an early age persist after maturity. A total of 60 female chicks were randomly assigned to one of two treatment groups: six pens with five chicks (brooded group) each were reared by a broody hen and six pens with five chicks (non‐brooded group) each were provided with an infrared heating lamp. We evaluated the persistent effects of broody hens by measures of behavior, physical condition and production at 9, 16, 35 and 55 weeks of age. The numbers of threatening, aggressive pecking, fighting and severe feather pecking behaviors were higher in non‐brooded than in brooded chickens (all P < 0.05). Egg production was lower in brooded than in non‐brooded chickens (P < 0.05), while the number of brooding chickens was higher in the brooded than in the non‐brooded group (P < 0.05). In conclusion, the presence of broody hens at an early stage of chicks' lives has a persistent effect on behavior. Although brooded chickens showed more brooding and lower egg production than non‐brooded chickens, feather pecking and aggressive interaction were decreased in brooded hens.     

Mechanisms responsible for reduced cardiotoxicity of mitoxantrone compared to doxorubicin examined in isolated guinea-pig heart preparations

We reported previously that doxorubicin, an anticancer agent that has an anthracycline structure, alters Ca2+ releasing and uptake mechanisms in the sarcoplasmic reticulum of myocardial cells. These effects of doxorubicin are apparently related to its cardiotoxicity. Mitoxantrone is a similar anticancer agent with an anthracenedion structure that has been shown to be significantly less cardiotoxic. In the present study, the effects of mitoxantrone on the functions of the sarcoplasmic reticulum were examined in isolated muscle preparations obtained from the guinea-pig heart. In electrically-stimulated left atrial muscle preparations, incubation in vitro for 4 hr with 30 or 100 microM mitoxantrone significantly prolonged the time to the peak of twitch tension, markedly increased the developed tension observed at lower stimulation frequencies, thereby attenuating the slope of positive force-frequency relationships, and increased the postrest contraction observed after a 60-sec quiescent period. In myocytes isolated from ventricular muscles, 30 microM mitoxantrone increased the peak and the size of intracellular Ca2+ concentrations ([Ca2+] i), and prolonged the time to peak [Ca2+]i. In skinned muscle fiber preparations obtained from the left ventricular muscle, 30 muM mitoxantrone significantly increased the caffeine-induced contraction without affecting the Ca2+ sensitivity of contractile proteins. These results suggest that mitoxantrone enhances Ca2+ release from the sarcoplasmic reticulum in isolated atrial muscle preparations obtained from the guinea-pig heart. Apparent enhancement of the sarcoplasmic reticulum functions, in contrast to anthracyclines that has been shown to suppress these functions, seems to explain the relative lack of marked cardiotoxicity of mitoxantrone.     

N,N'-Bis(2-chloroethyl)-N-nitrosourea (BCNU)-induced Apoptosis of Neural Progenitor Cells in the Developing Fetal Rat Brain

N,N'-bis(2-chloroethyl)-N-nitrosourea (BCNU) is one of the major drugs used in chemotherapy against malignant gliomas due to its effects, such as induction of bifunctional alkylation of DNA and formation of interstrand DNA cross-linkages, and induces cortical malformations in the fetal and neonatal rat brain. In this study, pregnant rats were treated with 7.5 mg/kg of BCNU on gestational day 13 (GD 13), and their fetuses were collected from 12 to 72 hours after BCNU treatment in order to examine the timecourses of morphological and immunohistochemical changes in neural progenitor cells in the developing brain. The number of pyknotic cells in the telencephalon peaked at 24 h and then gradually decreased until 72 h. The majority of these pyknotic cells were positive for cleaved caspase-3, a key executioner of apoptosis. The pyknotic cells showed the ultrastructural characteristics of apoptosis. The number of p53-positive cells began to increase prior to the appearance of apoptotic cells and p21-positive cells. The number of phosphorylated-histone H3-positive cells (mitotic cells) decreased from 24 to 36 h. The number of Iba1-positive cells (microglial cells) in the telencephalon increased from 12 to 48 h. These results suggest that BCNU induces p53-dependent apoptosis and reduces proliferative activity, resulting in reduction of the weight of the telencephalon and the thickness of the telencephalic wall in the fetal brain. This study will help to clarify the mechanisms of BCNU-induced fetal brain toxicity.     

Molecular cloning of canine membrane-anchored inhibitor of matrix metalloproteinase, RECK

The reversion-inducing cysteine-rich protein with Kazal motifs (RECK) gene is one of the endogenous matrix metalloproteinase (MMP) inhibitors. It was reported that decreased RECK expression closely correlated with tumor malignancy. We determined the cDNA sequence of the canine RECK gene. The cDNA sequence and deduced amino acid of canine RECK were 2,913 bases and 971 residues, respectively. The predicted amino acid sequence of the protein showed 95.5% and 91.9% homology with human and mouse RECK, respectively. RECK mRNA expression was analyzed in various canine tissues and tumor cell lines by quantitative RT-PCR. The highest RECK expression was detected in lung and testis. In comparison with the tissues, a remarkably low expression level was detected in tumor cell lines. In addition, the RECK gene was transfected in the canine transitional cell carcinoma, and its influence on cell proliferation, migration, and invasion was analyzed. The transfected RECK gene suppressed only canine tumor invasion. These results showed that RECK might play an important role in tumor malignancy in dogs as well as in other mammalians.     

Development of a SYBR green real-time polymerase chain reaction assay for quantitative detection of Babesia gibsoni (Asian genotype) DNA.

A real-time fluorogenic polymerase chain reaction (PCR) assay based on SYBR green that allows for sensitive, reproducible, and accurate quantification of Babesia gibsoni (Asian genotype). DNA from peripheral blood of infected dogs was developed. Standard curves were created by plotting the input amount of a standard template, constructed with plasmid DNA containing 182 base pairs (bp) of the p18 gene, against threshold cycle numbers. The curves showed a wide dynamic range (1,000,000-fold input) and high correlation values (>0.99). The PCR amplification efficacy of the standard template was similar to that of intact genomic DNA obtained from peripheral blood with B. gibsoni infection. The detection limit of the assay was 9 parasites/microl of blood with B. gibsoni infection. The intra-assay and interassay coefficients of variation of the threshold cycles ranged from 0.70% to 1.89% and from 1.18% to 1.92%, respectively. This assay system was found to be reproducible and accurate for the quantification of parasite DNA in experimentally infected dogs and far more sensitive than traditional microscopic examination.     

Molecular Detection of Avian Pathogens in Poultry Red Mite (Dermanyssus gallinae) Collected in Chicken Farms

Poultry red mite (PRM, Dermanyssus gallinae) is a blood-sucking ectoparasite as well as a possible vector of several avian pathogens. In this study, to define the role of PRM in the prevalence of avian infectious agents, we used polymerase chain reaction (PCR) to check for the presence of seven pathogens: Avipox virus (APV), Fowl Adenovirus (FAdV), Marek’s disease virus (MDV), Erysipelothrix rhusiopathiae (ER), Salmonella enterica (SE), Mycoplasma synoviae (MS) and Mycoplasma gallisepticum (MG). A total of 159 PRM samples collected between 2004 and 2012 from 142 chicken farms in 38 prefectures in Japan were examined. APV DNA was detected in 22 samples (13.8%), 19 of which were wild-type APV. 16S ribosomal RNA (16S rRNA) of MS was detected in 15 samples (9.4%), and the mgc2 gene of MG was detected in 2 samples (1.3%). Eight of 15 MS 16S rRNA sequences differed from the vaccine sequence, indicating they were wild-type strains, while both of the MG mgc2 gene sequences detected were identical to the vaccine sequences. Of these avian pathogen-positive mite samples, three were positive for both wild-types of APV and MS. On the other hand, the DNAs of ER, SE, FAdV and MDV were not detected in any samples. These findings indicated that PRM can harbor the wild-type pathogens and might play a role as a vector in spreading these diseases in farms.     

Impacts of pellet pH on nitrous oxide emission rates from cattle manure compost pellets

Previous reports indicated that the emission of nitrous oxide (N₂O) when manure compost pellets (MCP) were applied to soil was greater than when ordinary manure compost or inorganic fertilizer was applied, but that applying pellets of nitrogen-enriched manure compost, a by-product of deodorizing manure during composting, resulted in N₂O emission rates less than those from MCPs. To investigate the mechanism by which N₂O emission rates and cumulative emissions were reduced in nitrogen-enriched manure composts pellets (N+MCP), we studied the impact of pellet pH on N₂O emission, because pH is different between MCP (pH 8.6) and N+MCP (pH 5.3). In an incubation experiment, the pH of pellets was adjusted to five levels (5.3, 6.0, 7.0, 8.0 and 8.6) with acid or alkaline solutions, and the pellets were incubated without soil in a beaker at 30°C for 90 d (MCP) or 42 d (N+MCP). A large peak in N₂O emission rate was observed soon after beginning the incubation (within 1–3 d) in the neutral and alkaline treatments for both MCP and N+MCP, and these peaks corresponded to a rise in the pellet nitrite contents. Thus, this N₂O emission peak might have been generated by the denitrification of nitrite in the pellets. In the acid treatments of MCP, the N₂O emission was distributed more in the later incubation period (14–90 d), when the reduction of nitrate in MCP occurred. This led to a significant increase in cumulative N₂O emission as compared with the alkaline treatments for MCP. Regarding the mechanism by which N₂O emission was reduced in N+MCP, although larger cumulative N₂O emission rates in the earlier stage (0–14 d for MCP and 0–7 d for N+MCP) were observed when the pellet pH was adjusted close to 7.0, lowering the pH of MCP to 5.3 (the pH of N+MCP) did not demonstrate a significant decrease in cumulative N₂O emission as compared with the original pH treatment (pH 8.6). These results indicate that pellet pH might not relate directly to the mechanism by which N₂O emission was reduced in N+MCP.     

Protein metabolism in rice seedling

The relationship between protein synthesis and degradation in germinating rice seed were studied with protein synthetic inhibitors. Both DNP and 8-AG inhibited the degradation of glutelin, the major storage protein in rice seed, while the inhibitors had no direct effect in the activity of rice seed proteinllse in vitro. The prevention can be partly ascribed to the Inhibition of proteinase synthesis because the inhibitors depressed the increase in proteinase activity during germination. When DNP treatment was started at the onset of germination, the degradation of glutelin in the endosperm was seriously inhibited and the endosperm remained rigid over 9 days of incubation at 30°C. In contrast, the inhibition was less efficient clent when the treatment was started in the later stage. It is suggested that the degradation of the storage protein in rice seed depends on the synthetic process of the hydrolytic enzymes which increase during germination. disintegrate the compartmentation of the endosperm and allow the storage proteins to come in contact with the existing proteinases     

Annual reproductive cycle of the greasyback shrimp Metapenaeus ensis in Ise Bay,Japan

The greasyback shrimp Metapenaeus ensis is widely distributed along the coast of India and the West Pacific where it is an important fisheries species. We have examined seasonal changes in ovarian development, spermatogenesis, and mating of Me. ensis in histological studies and by external observations on specimens collected in Ise Bay, its northernmost habitat. Ovaries were found to be previtellogenic from February to May, with the first signs of development being the accumulation of yolk in oocytes in late May. Ovarian shadow ratios were high during the period late July to mid-September. The formation of cortical rods in the peripheries of oocytes and germinal vesicle breakdown were observed in ovaries from late June to September. Male shrimps had sperm in the testes during the period early June to early October, and female shrimps had spermatophores in spermatheca after early July. In late July, some post-spawn female shrimps had exogenous vitellogenic oocytes in their ovaries, indicating that ovarian development had been repeated in preparation for the next spawning. Ovarian shadow ratios, which were positively correlated with gonadosomatic indices and ovarian development, seem to be a useful marker to determine ovarian development. Our results indicate that mating in Me. ensis started in early July and that the spawning season ranged from July to September with more than two cycles of spawning in Ise Bay.