Real-time PCR genotyping assay for feline erythrocyte pyruvate kinase deficiency and mutant allele frequency in purebred cats in Japan

Erythrocyte pyruvate kinase (PK) deficiency is an inherited glycolytic erythroenzymopathy caused by mutations of the PKLR gene. A causative mutation of the feline PKLR gene was originally identified in Abyssinian and Somali cats in the U.S.A. In the present study, a TaqMan probe-based real-time PCR genotyping assay was developed and evaluated for rapid genotyping and large-scale screening for this mutation. Furthermore, a genotyping survey was carried out in a population of four popular purebred cats in Japan to determine the current mutant allele frequency. The assay clearly displayed all genotypes of feline PK deficiency, indicating its suitability for large-scale survey as well as diagnosis. The survey demonstrated that the mutant allele frequency in Abyssinian and Somali cats was high enough to warrant measures to control and prevent the disease. The mutant allele frequency was relatively low in Bengal and American Shorthair cats; however, the testing should still be carried out to prevent the spread of the disease. In addition, PK deficiency should always be considered in the differential diagnosis of anemia in purebred cats in Japan as well as worldwide.     

Tooth brushing inhibits oral bacteria in dogs

In this study, scaling, polishing and daily tooth brushing were performed in 20 beagle dogs, and the number of oral bacteria was determined using a bacterial counter. The dogs were randomized into the scaling (S), scaling + polishing (SP), scaling + tooth daily brushing (SB) and scaling + polishing + tooth daily brushing (SPB) groups. Samples were collected from the buccal surface of the maxillary fourth premolars of the dogs immediately after scaling and every week thereafter from weeks 1 to 8. Throughout the study, the number of bacteria was significantly lower in the SB and SPB groups compared with the S group. The findings suggest that daily tooth brushing inhibited oral bacterial growth in the dogs.     

The status of atmospheric concentrations of ammonia in an intensive dairy farming area in central Japan

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Silicon availability in 36 commercial nursery bed soils was evaluated by four methods the phosphate buffer (pH 6.2, 40 mmol L⁻¹), incubation, supernatant and acetate buffer (pH 4.0, 1 mol L⁻¹) Methods. The influence of silicon availability in the nursery bed soils on the silicon uptake of rice Oryza sativa L. cv. Hitomebore seedlings and the effect of silicon fertilizer application were examined in a glass house in 2002.
The results revealed that the best correlation between silicon content in rice seedlings and available silicon in soils was obtained with the phosphate buffer-solution method ( r  = 0.86). More precise evaluation of available silicon was achieved by grouping soils based on these phosphate absorption coefficients (PAC). The correlation coefficients between silicon content in rice seedlings and available silicon in soils were 0.92 and 0.72 for volcanic soils (PAC > 1500) and non volcanic soils (PAC < 1500), respectively.
We concluded that the phosphate buffer method is the most easily adjusted method for estimation of silicon availability in nursery bed soils, and silicon fertilizers should be applied when silicon availability in non-volcanic nursery bed soils goes below 200 mg kg⁻¹, whereas the level is less than 350 mg kg⁻¹ in volcanic soils.     

Evaluation of a Collagenase Generated Osteoarthritis Biomarker in Naturally Occurring Canine Cruciate Disease

Objective— To determine the clinical value of a novel osteoarthritis (OA) biomarker in detecting canine cruciate disease.
Study Design— Cross sectional clinical study.
Animals— Dogs (n=22) with cranial cruciate ligament (CCL) rupture and 12 control dogs.
Methods— Concentrations of collagenase-generated cleavage epitope of type II collagen (Col2-3/4C_{long mono}, or C2C) in serum, urine, and joint fluid were compared between a group of dogs with CCL rupture and a control group. Correlation of C2C concentrations to the clinical stage of stifle OA was also evaluated.
Results— There were no significant differences in C2C concentrations in serum, urine, and joint fluid between groups ( P >.05). Subjective scores of lameness, joint effusion, osteophytosis were significantly more severe in the CCL rupture group compared with the control group ( P <.05). There was no significant correlation of C2C concentrations with clinical stage of stifle OA ( P >.05).
Conclusion— This OA biomarker did not detect pathology associated with CCL rupture. Our results suggest that collagenase-specific degradation of type II collagen in articular cartilage may not be involved in the early stage of naturally occurring canine cruciate disease, and that pathology associated with naturally occurring CCL rupture is different from that of experimental OA model.
Clinical Relevance— C2C is not clinically useful in detecting CCL rupture in dogs.     

Formation of zinc protoporphyrin IX in Parma-like ham without nitrate or nitrite

Zinc protoporphyrin IX (ZPP) is a characteristic red pigment in meat products that are manufactured without the addition of a curing agent such as nitrate or nitrite. To examine the effects of impurities such as mineral components in sea salt on the formation of ZPP, we manufactured Parmatype dry-cured hams that were salted with refined salt or sea salt and examined the involvement of oxidation-reduction potential (ORP) in the formation of ZPP. The content of ZPP was increased drastically after 40 weeks. Microscopic observation showed strong fluorescence caused by ZPP muscle fiber after 40 weeks. Conversely, heme content varied considerably during processing. ORP increased during processing. However, there was no obvious difference between ham salted with refined salt and that salted with sea salt. Therefore, it was concluded that impurities in sea salt were not involved in the formation of ZPP.     

Effects of maize (Zea mays L.) silage feeding on dry matter intake and milk production of dairy buffalo and cattle in Tarai, Nepal

To identify the effects of whole crop maize silage (MS) as a substitute for rice straw (RS) on feed intake and milk production of mid-late lactating buffalo and cattle in Tarai, Nepal, eight Murrah and eight Jersey-Hariana were fed the basal diet, RS ( ad libitum ) with concentrate (0.68% of bodyweight [BW] on a dry matter [DM] basis). A 4 × 4 Latin square design experiment was conducted in each animal species with graded levels of MS substitution for RS (0%, T1; 33%, T2; 67%, T3 and 100%, T4). The MS had higher digestibility and total digestible nutrient (TDN) than RS. The DM intake per BW of the both species was highest in T3. The substitution of MS for RS increased the crude protein intake and the TDN intake in the both species. Although the buffalo showed the highest milking performance in T4, the cattle showed no significant differences in their milking performance among the treatments. The substitution of MS for RS improved the feed intake and milk production in the buffalo. On the other hand, the milk yield was not raised in the cattle, though the feed intake was increased by the substitution.     

Psycho-educational Horseback Riding to Facilitate Communication Ability of Children with Pervasive Developmental Disorders

In this study, we applied a novel psycho-educational horseback riding (PEHR) program to the treatment of four Japanese children with pervasive developmental disorders (PDD) in order to facilitate the acquisition of verbal and nonverbal communication skills. The behavioral changes in each child were evaluated using a psychological and behavioral scale. The scale for evaluating the effect of Human-Equips-Interaction on Mental activity (HEIM scale) was designed to assess the behavioral improvement of children based on the following 10 items: Human relationships, Imitation, Emotional expression, Sudden physical movement, Fixative behavior, Adaptation to change, Visual response, Fear or nervousness, and Verbal and nonverbal communication. After taking part in the PEHR program for several months, all subjects showed remarkably improved HEIM scores and marked improvements were observed in eye contact with others (instructors, side walkers, and leaders) in the riding area. A statistical difference was found in items 1, 2, 3, 6, 7, 8, and 9. However, no statistical difference was found in items 4, 5, and 10. As the program progressed, the children showed enhanced verbal and nonverbal communication skills, and became more expressive in their emotional and empathetic interaction with their parents. These observations suggest that the normal functioning of pleasurable emotions and empathy may facilitate further improvements in joint attention, imitation and empathy, and may result in successful verbal expression by PDD children. Therefore, horseback riding can play a very important role in the psycho-educational support required for the communication ability of PDD children.     

Water velocity in artificial habitats of the Japanese crayfish Cambaroidesjaponicus

ABSTRACT:   The appropriate water velocity in artificial burrows for theJapanese crayfish species Cambaroides japonicus was studied.An experimental system of two artificial burrows (burrows A and B)of the same size was set in a large tank. The velocity of burrowA was 0 cm/s and that of burrow B was varied andset at 0 cm/s, 5 cm/s, 10 cm/s,20 cm/s and 30 cm/s. The selectionof these two burrows by C. japonicus was observed. No animalsselected burrow B significantly more than burrow A above 10 cm/s.At 20 cm/s, some crayfishes were swept away andcould not return to burrows because of the high water velocity.At 30 cm/s, most animals were swept away. We concludethat the appropriate water velocity for the suitability of artificialburrows and the immediate foraging area adjacent to the burrowsshould be as low as 5 cm/s.     

Improvement of the emulsifying properties of porcine sarcoplasmic proteins with a low-molecular weight surfactant by the two-step emulsification method

The emulsifying properties of porcine sarcoplasmic protein (SP) with or without low-molecular weight surfactants were examined. The emulsifying activity (EA) of SP was weaker than that of sodium caseinate (CA), blood plasma (BP) and whey protein isolates at a protein concentration of 0.5–3%. Sodium chloride decreased the EA of SP. The EA of SP was about one quarter that of CA in the presence of 3% sodium chloride. As homogenization time increased, the EAs of SP and BP decreased. The EA of SP was not influenced by sucrose stearate ester (SE) with a hydrophilic-lipophilic balance (HLB) value of 16 (SE-16) but was decreased by those with HLB values of 5 and 1. Low-molecular weight surfactants such as SE-16, Tween 80 and sodium cholate improved the EA of SP with the two-step emulsification method: corn oil was previously emulsified with the surfactant before addition of SP and homogenization. In the case of the addition of SE-16 as primary emulsifier, the EA was about twice as high as that of SP alone. The same effect was shown in the emulsions formed by CA and soy protein isolate. These results indicate that the emulsifying properties of meat protein would be improved by the addition of a low-molecular weight surfactant in a two-step emulsification.