Development and characterization of monoclonal antibodies against canine trypsin

Canine cationic trypsin was purified by salting-out, gel filtration and affinity chromatography. Purity was confirmed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The molecular weight was ca. 28 kDa by SDS-PAGE.

Thirty hybridomas were obtained which produced mAb to canine cationic trypsin by the cell fusion technique. Twenty-two of these recognized cationic trypsin only, while eight hybridomas recognized both cationic and anionic trypsin. Several of the anti-canine cationic trypsin mAb were purified by salting-out and DEAE ion-change chromatography using ascites fluid of immunized BALB/c mice. The mAb proved to have very high specificity to canine cationic trypsin as shown by immunoblotting and it may be possible to use them to develop clinical assays.     

Whole-Cell Fatty Acid Composition to Characterize and Differentiate Isolates of Rhizoctonia Species Associated with Turfgrass Diseases in Japan

Cellular fatty acids were analyzed to characterize and differentiate 34 isolates of Rhizoctonia species representing binucleate Rhizoctonia AG-D (I), AG-D (II), R. solani AG 2-2 IIIB, AG 2-2 LP, R. circinata var. circinata and var. oryzae associated with turfgrass diseases in Japan. Myristic, pentadecanoic, palmitic, palmitoleic, stearic, oleic, linoleic and linolenic acids were consistently present in varying quantities in all isolates. Heptadecanoic and 9-heptadecenoic acids were present in isolates of Rhizoctonia AG-D (I), AG-D (II), R. solani AG 2-2 IIIB and AG 2-2 LP but not in isolates of R. circinata var. circinata and var. oryzae. Palmitic, oleic and linoleic acids were the major fatty acids found, constituting 88.30-98.37% of the whole-cell fatty acid content. The remaining fatty acids were present in smaller amounts. Isolates within a single group were closely clustered, whereas isolates from different groups were clearly distinguishable based on average linkage cluster analysis of cellular fatty acids. Principal component analysis, based on all fatty acids detected, confirmed the distinct separation of isolates representing the six groups of Rhizoctonia species obtained from turfgrasses. These results suggested that fatty acid analysis is useful for the characterization and differentiation of isolates of Rhizoctonia species associated with turfgrass diseases. Received 21 May 2001/ Accepted in revised form 28 September 2001     

Performance of Bangladesh indigenous rice in a weed infested field and separation of allelopathy from resource competition

This study aimed to identify the potential allelopathic indigenous rice (Oryza sativa L. ssp. indica) varieties from Bangladesh using a performance study in a weed‐infested field and to assess the extent of allelopathic interference relative to resource competition in a glasshouse experiment. Six varieties – namely, “Boterswar,” “Goria,” “Biron” and “Kartiksail” as the most allelopathic, “Hashikolmi” as weakly allelopathic and “Holoi” as nonallelopathic – were raised following a nonweed control method. The infestation levels of weed species were calculated using Simpson's Diversity Index (SDI), which ranged from 0.2 to 0.56. However, a significant correlation coefficient (0.87, P < 0.001) was obtained from these field data compared with the root inhibition percentage from the laboratory bioassay, and the “Boterswar” variety was the most allelopathic. The interactions between the allelopathic variety “Boterswar,” weakly allelopathic variety “Hashikolmi” and Echinochloa oryzicola via a target (rice)‐adjacent (E. oryzicola) cogrowth culture were determined in a hydroponic arrangement. The relative competitive intensity (RCI) and the relative neighbor effect (RNE) values showed that the crop–weed interaction was facilitation for “Boterswar” and competition for “Hashikolmi” and E. oryzicola in rice/E. oryzicola cogrowth cultures. The allelopathic effects of “Boterswar” were much higher than the resource competition in rice/E. oryzicola cogrowth cultures. The converse was observed for “Hashikolmi.” Moreover, the mineral content of E. oryzicola was severely affected by “Boterswar”/E. oryzicola cogrowth cultures’ exudate solution. Therefore, the allelopathic potential of “Boterswar” variety might be useful for developing the weed‐suppressing capacity of rice, which will likely have a significant influence on paddy weed control.     

Identification of the core motif of the BRCA2 C-terminal RAD51-binding domain by comparing canine and human BRCA2

Mammary tumors are the most common tumors in women and non-spayed female dogs. One of the reasons for mammary tumors is mutations of the tumor suppressor gene, BRCA2. BRCA2 participates in homologous recombination repair by interacting with the RAD51 recombinase. BRCA2 has two RAD51-binding domains, consisting of BRC repeats and the C-terminal RAD51-binding domain, respectively. Although several studies have addressed the function of the C-terminal RAD51-binding domain of human BRCA2, the amino acid sequences required for the RAD51-interaction activity remain unclear. In this study, the C-terminal RAD51-binding domains of canine and human BRCA2 were compared; the canine domain displayed a weaker interaction with RAD51. This difference was attributed to the C-terminal portion of the domain via a comparison between canine and human domains. Furthermore, peptides shorter than those previously reported displayed RAD51-interacting activity, and a core motif of this domain consisting of 25 amino acids was identified. Since a mutation (S3323N) was reported in the core motif of this domain, the effect of this mutation was evaluated. The mutant exhibited similar RAD51-binding activity as that of the wild-type protein, suggesting that the mutation was functionally neutral. These data suggested that the C-terminal portion of the BRCA2 C-terminal RAD51-binding domain influenced its RAD51-interaction activity, and a minimum core motif of 25 amino acids was identified in this domain. These data may help clarify BRCA2 function, as well as the tumorigenic effects of BRCA2 mutation.     

Identification of the core rumen bacterial taxa and their population dynamics during the fattening period in Japanese Black cattle

The rumen microbiota comprises a vast range of bacterial taxa, which may affect the production of high-quality meat in Japanese Black cattle. The aim of this study was to identify core rumen microbiota in rumen fluid samples collected from 74 Japanese Black cattle raised under different dietary conditions using 16S rRNA gene amplicon sequencing. In the rumen of fattening Japanese Black cattle, 10 bacterial taxa, showing >1% average relative abundance and >95% prevalence, irrespective of the dietary conditions and the fattening periods, were identified as the core rumen bacterial taxa, which accounted for approximately 80% of the rumen microbiota in Japanese Black cattle. Additionally, population dynamics of the core rumen bacterial taxa revealed two distinct patterns: Prevotella spp. and unclassified Bacteroidales decreased in the mid-fattening period, whereas unclassified Clostridiales, unclassified Ruminococcaceae, Ruminococcus spp., and unclassified Christensenellaceae increased during the same period. Therefore, the present study reports the wide distribution of the core rumen bacterial taxa in Japanese Black cattle, and the complementary nature of the population dynamics of these core taxa, which may ensure stable rumen fermentation during the fattening period.     

A review of okara (soybean curd residue) utilization as animal feed: Nutritive value and animal performance aspects

Year by year, huge quantities of by-products are generated during the manufacturing process of soybean-based products. Okara is one of the by-products, and it is an insoluble portion of the soybean. It consists of high moisture (8.4–22.9%); on dry matter basis, it contains high metabolizable energy (9.0–14.2 MJ/kg) and other components that include crude protein (20.9–39.1%), crude fiber (12.2–61.3%), crude fat (4.9–21.5%), and ash (3.4–5.3%). Fermentation of okara improves its nutritional quality and reduces its anti-nutrient contents. Due to animals' palatability, okara can be used to replace the soybean meal/concentrate feed partially or completely in ruminant's diet and partially in nonruminant's diet. Okara feeding does not depress the intake, digestibility, growth, milk production, blood metabolic profiles, and meat quality of animals. However, this by-product decays quickly due to its high moisture content, and its heavy weight and sticky nature make it difficult to process and expensive to dry using conventional methods. This paper thoroughly summarizes the utilization of okara as animal feed in the cause of developing a general guideline with favorable levels of inclusion in the diets of animals for its exploitation and valorization. This review will encourage further research to develop eco-friendly and value added feed for animals.     

Genetic analysis for sow stayability at different parities in purebred Landrace and Large White pigs

Genetic parameters for sow stayability were estimated from farrowing records of 10,295 Landrace sows and 8192 Large White sows. The record for sow stayability from parity k to parity k + 1 (k = 1, …, 6) was 0 when a sow had a farrowing record at parity k but not at parity k + 1, and 1 when a sow had both records. Heritability was estimated by using single-trait linear and threshold animal models. Genetic correlations among parities were estimated by using two-trait linear–linear and single-trait random regression linear animal models. Genetic correlations with litter traits at birth were estimated by using a two-trait linear–linear animal model. Heritability estimates by linear model analysis were low (0.065–0.119 in Landrace & 0.061–0.157 in Large White); those by threshold model analysis were higher (0.136–0.200 & 0.110–0.283). Genetic correlations among parities differed between breeds and models. Genetic correlation between sow stayability and number born alive was positive in many cases, implying that selection for number born alive does not reduce sow stayability. The results seem to be affected by decisions on culling made by farmers.     

Changes in rat uterine and cervical phospholipase A2 activity following progesterone agonist or antagonist administration at term

Our previous study revealed that a fall in plasma progesterone (P(4)) level was associated with a transient increase in cytosolic phospholipase A(2) (PLA(2)) activity and prostaglandin F(2)alpha level in the rat uterus and cervix during natural parturition. This study determined the changes in the PLA(2) activities during modulated occurrence of delivery by P(4) antagonist or agonist late in pregnancy. In rats undergoing P(4) antagonist-induced preterm delivery, the PLA(2) activities of both uterine and cervical cytosol significantly decreased 12 h after the challenge and tended to be attenuated within 72 h. The plasma P(4) level altered in a similar pattern. Blockade of delivery by chronic treatment with P(4) agonist was not associated with changes in uterine PLA(2) activity compared with that in normally delivering rats, although there was a persistent rise in cervical PLA(2) activity. The obtained data indicates that the PLA(2) activities in rat uterine and cervical cytosol are not regulated solely by P(4) and that delivery can occur without activation of this enzyme.     

Direct effects of prolactin on adrenal steroid release in male Hatano high-avoidance (HAA) rats may be mediated through Janus kinase 2 (Jak2) activity

Prolactin (PRL) has been proposed to directly stimulate corticosterone release. However, the role of PRL on adrenocortical function in male HAA rats has not been fully clarified. The aim of this study was to investigate the effect of PRL on the secretion of corticosterone and progesterone using an in vitro cell culture system in male rats. Administration of PRL (10(-7) and 10(-6) M) resulted in dose-dependent increases in corticosterone and progesterone release. Cotreatment with PRL produced an increase in the stimulatory effects of ACTH-induced corticosterone and progesterone secretion. However, the PRL-induced corticosterone and progesterone releases were significantly reduced by treatment with AG490, a specific Janus kinase 2 (Jak2) inhibitor. In addition, administration of AG490 blunted the significant inhibition of ACTH-induced corticosterone and progesterone secretion by PRL. These results demonstrated that PRL could act directly on the adrenal gland to drive corticosterone and progesterone secretion in male rats. Additionally, the results emphasize that PRL stimulation of adrenal steroid release may be mediated through Jak2 activity.