Relationship between transportation stress and polymorphonuclear cell functions of bottlenose dolphins, Tursiops truncatus

Dolphins in a captive environment are exposed to various kinds of stresses. Handling and transportation are stressful events for terrestrial mammals, and such stress may affect immune system function and increase susceptibility to infectious diseases. The same phenomenon could occur in dolphins, however, few studies have reported this in dolphins. The objective of this study was to evaluate the relationship between stress and polymorphonuclear (PMN) cell function of dolphins during transportation. Four bottlenose dolphins (Tursiops truncatus) were transported for 6 hr by truck. Serum cortisol levels, leukograms, phagocytosis, and superoxide production of PMN cells were evaluated during handling and transportation compared to resting values. The mean serum cortisol level was significantly increased during handling and transportation (p<0.05) when compared with the resting values. White blood cell (WBC) counts, eosinophil counts, phagocytosis, and superoxide production of PMN cells during handling and transportation stages decreased significantly in comparison with the resting stage (p<0.05). The concentration of serum cortisol was significantly correlated with the results of the WBC counts, eosinophil counts, superoxide production, and phagocytosis (p<0.01, p<0.05, p<0.05, and p<0.001, respectively). The present results indicate that handling and transportation are stressful events for dolphins and could affect their PMN cell functions, thereby leading to the impairment of the immune system.     

Suppurative meningoencephalitis and perineuritis caused by Streptococcus gallolyticus in a Japanese Black calf

A 179-day-old calf, which was weak and stunted, showed neurological signs and was euthanized. Postmortem examination revealed extensive and severe cloudy area in the meninges, and pleural pneumonia. Gram-positive cocci were isolated from systemic organs. Biochemical and 16S rRNA gene sequence analyses identified the isolate as Streptococcus gallolyticus, and its subspecies was suggested to be gallolyticus (SGG). The isolate was classified as a novel sequence type (ST115) by the multilocus sequence typing scheme for SGG and showed susceptibility to penicillin, ampicillin, amoxicillin, florfenicol, sulfamethoxazole-trimethoprim, and chloramphenicol. Histopathologically, suppurative meningoencephalitis and perineuritis were detected. As SGG has been isolated solely from a cow with mastitis in Japan, this is the first SGG infection in a calf with suppurative meningoencephalitis and perineuritis in this country.     

Artificially induced tetraploid masu salmon have the ability to form primordial germ cells

Diploid gametes generated with tetraploid animals are a stepping stone to improving chromosome manipulation techniques. However, artificially induced tetraploid individuals generally die soon after hatching. Diploid gametes could be induced by in vivo cultures of tetraploid primordial germ cells (PGCs) through germ-line chimera. In the present study, characteristics of PGCs were studied in inviable tetraploid masu salmon, Oncorhynchus masou. Histological observation of tetraploid embryos revealed that the same or smaller numbers of PGCs were observed and they migrate into the genital ridges as did diploid PGCs during gonadogenesis. By whole-mount in situ hybridization using vasa messenger RNA (mRNA), 4–35 vasa-positive signals were detected in a pair of genital ridges of tetraploids. By cytological observation of genital ridge cell suspensions, several large round cells were observed, some of which extended pseudopodia. They also contained large nuclei and round granules in their cytoplasm, characteristics of PGCs. As the results suggest that inviable artificial tetraploids have PGCs, we expect to achieve diploid gamete production through surrogate propagation and tetraploid fish production.     

Comparative intestinal absorption of amino acids in rainbow trout (Oncorhynchus mykiss), totoaba (Totoaba macdonaldi) and Pacific bluefin tuna (Thunnus orientalis)

The kinetics of amino acid absorption in the proximal section of three fish species were studied using an everted intestine technique and a pancreatic digest of casein (Tryptone), as the model amino acid mixture. Fresh intestine of rainbow trout (Oncorhynchus mykiss) was used to set up the experimental system, and results were compared with those obtained using intestines from totoaba (Totoaba macdonaldi) and bluefin tuna fish (Thunnus orientalis). The kinetics of amino acid absorption was sigmoidal for all amino acids and for each species of fish tested. In general, specific absorptions of essential amino acids were higher than those of non‐essential ones. No correlation between the concentration of amino acids in the Tryptone solution and the corresponding absorptions was found. The maximum specific absorption rates of all amino acids for trout were 10 times higher than those determined for totoaba and bluefin tuna. The relative amounts of the different amino acids preferentially absorbed in all three species were different. Results obtained from the everted technique may be applicable to the design of formulated diets for large fish species with commercial value.     

Effect of presence of rumen protozoa on degradation of cell wall constituents in gastrointestinal tract of cattle

The cell wall constituents of feces from three faunated and three defaunated (without ruminal ciliate protozoa) cattle fed on a Sudangrass hay and concentrate mixture (8:5) were analyzed. There was little difference in digestibility of dry matter between the faunated and defaunated cattle. Analysis of the fecal sugar residues revealed that the digestibilities of arabinose and galactose, derived from pectic and hemicellulosic substances located within the compound middle lamella, were higher in the defaunated cattle than the faunated cattle (P < 0.05), whereas the digestibilities of glucose and xylose, derived mainly from cellulose and xylan, were unchanged by the removal of protozoa. The digestibility of lignin was not different between the faunated and defaunated cattle, but those of mannose and p‐coumaric acid were lower in the defaunated than in the faunated animals (P < 0.05). The ratio of primary cell wall to secondary cell wall in fecal plant materials was lower for the defaunated than for the faunated cattle. The results in this study suggested that the defaunation enhanced the microbial degradation of the thin cell walls, but depressed the degradation of developed cell walls.     

Evaluating environmental impacts of the Japanese beef cow–calf system by the life cycle assessment method

The objectives of this study were to evaluate the environmental impacts of a beef cow–calf system using a life cycle assessment (LCA) method and to investigate the effects of scenarios to reduce environmental impacts on the LCA results. The functional unit was defined as one marketed beef calf, and the processes associated with the cow–calf life cycle, such as feed production, feed transport, animal management, the biological activity of the animal and the treatment of cattle waste were included in the system boundary. The present results showed that the total contributions of one beef calf throughout its life cycle to global warming, acidification, eutrophication and energy consumption were 4550 kg of CO₂ equivalents, 40.1 kg of SO₂ equivalents, 7.0 kg of phosphate (PO₄) equivalents and 16.1 GJ, respectively. The contribution of each process to the total environmental impact in each environmental impact category showed a similar tendency to the contribution of each process in each environmental category reported in the case of the beef fattening system as a whole. The results from this analysis showed that shortening calving intervals by 1 month reduced environmental impacts by 5.7–5.8% in all the environmental impact categories examined in the current study, and increasing the number of calves per cow also reduced environmental impacts in all the categories, although the effects were smaller.     

The addition of calcium ion chelator,EGTA to thawing solution improves fertilizing ability in frozen–thawed boar sperm

In our previous study, seminal plasma effectively suppressed the induction of sperm to capacitation‐like status and acrosome loss during the thawing process. However, because boar seminal plasma is contaminated with various kinds of bacteria and/or viruses, it is necessary to develop a thawing solution without animal‐derived materials. In this study, we focused on the increase of intracellular Ca²⁺ ([Ca²⁺]_i) in sperm after thawing and the negative effects of sperm qualities. After thawing, the fluorescent intensity of [Ca²⁺]_i indicator, Fluo‐3/AM, and the level of phosphorylated tyrosine residue of protein were increased in the sperm. Next, we investigated whether the addition of Ca²⁺ chelators (ethylenediaminetetraacetic acid (EDTA) and ethyleneglycoltetraacetic acid (EGTA)) improved post‐thawed sperm motility. When the frozen–thawed sperm were treated with 6 mmol/L EDTA + 6 mmol/L EGTA, sperm motility was significantly increased as compared with control (6 mmol/L EDTA alone) at all incubation periods (P < 0.05). The combinational treatment significantly suppressed the elevation of [Ca²⁺]_i and the tyrosine phosphorylation, which improved the acrosomal status and fertilizing ability in vitro. Furthermore, when the thawing method was applied for artificial insemination, the fertilization rate was significantly higher than control (P < 0.05, 33% vs. 82%). Thus, we conclude that the addition of EDTA + EGTA to thawing solution is a beneficial tool for artificial insemination using frozen–thawed boar sperm.     

Oxidative dimers produced from protocatechuic and gallic esters in the DPPH radical scavenging reaction

DPPH (2,2-diphenyl-1-picrylhydrazyl) radical scavenging reactions of protocatechuic and gallic acids, and their methyl esters, have been investigated by NMR. In acetone, methyl protocatechuate was gradually converted to a Diels-Alder adduct of two molecules of the intermediate quinone in the reaction with DPPH radical, whereas methyl gallate rapidly gave a symmetrical dimer via a putative quinone precursor. Both dimers are rather unstable and their structures have been deduced by in situ NMR measurements of the reaction mixtures. Gallic acid also gave a corresponding symmetrical dimer in the same reaction as methyl gallate, although protocatechuquinone produced from protocatechuic acid did not yield a Diels-Alder adduct, unlike its methyl ester. Interestingly, these dimer formations were not observed in methanol solution.