Three-dimensional reconstruction of intramuscular collagen networks of bovine muscle: A demonstration by an immunohistochemical/confocal laser-scanning microscopic method

We undertook a three‐dimensional reconstruction of intramuscular collagen networks of bovine muscle using an immunohistochemical/confocal laser‐scanning microscopic method. By immunohistochemical staining, type I and III collagens were observed mainly in the perimysium, while type IV collagen was observed in the endomysium. On the other hand, type V and VI collagens were observed in both the perimysium and endomysium. By confocal laser‐scanning microscopy, the collagen observed in the perimysium was three‐dimensionally reconstructed as plate‐shaped layers whereas the collagen observed in the endomysium surrounded myofibers. The three‐dimensionally reconstructed observations using immunohistochemical/confocal laser‐scanning microscopic method is useful for investigating collagen networks in muscle.     

Analysis of weed vegetation of no-tillage upland fields based on the multiplied dominance ratio

The objective of the present study was to clarify the characteristics of weed vegetation in no-tillage upland fields in Japan. Vegetation surveys were conducted in two stages and the obtained data were analyzed based on the multiplied dominance ratio. The first was a two-year survey of a no-tillage field and a tilled field where no-tillage or tillage farming had been carried out for more than 10 years. The no-tillage field surveyed exhibited a different seasonal succession from the tilled field. Summer annuals dominated the tilled field in summer, and winter annuals dominated in spring. Perennials occupied the no-tillage field in spring; summer annuals also dominated in summer. The second survey was of 19 and 22 fields, including some no-tillage fields, in the summer of 2000 and the spring of 2001. The vegetation in both summer and spring was classified into three vegetation types, principally dependent on the no-tillage periods: types I and II consisted mainly of tilled and under 3-year-old no-tillage fields, and type III comprised no-tillage fields three years old or over. Type II vegetation was characterized primarily by the presence of biennials and perennials of Asteraceae, and type III by perennials, such as sweet vernalgrass ( Anthoxanthum odoratum ), in addition to the same species as type II. Summer annuals such as southern crabgrass ( Digitaria ciliaris ) were abundant in many fields in summer regardless of the vegetation types.     

Inoculation of Lupinus luteus with White Root Rot Fungus, Rosellinia necatrix, to Estimate Virulence

An inoculation method using Lupinus luteus was developed for estimating virulence of isolates of the white root rot fungus, Rosellinia necatrix Prillieux. Fungal cultures grown on pieces of mulberry twigs were placed in contact with the hypocotyl of 3-week-old seedlings growing in pots of soil. Disease development was uniform and reproducible in repeated experiments. Of 24 isolates with double-stranded RNA, eight were weakly virulent. This method is useful throughout the year for estimating virulence of many isolates of the fungus and for screening for hypovirulent isolates. Received 2 March 2001/ Accepted in revised form 25 May 2001     

Characterization of two microsatellites in chicken monoamine oxidase A

We surveyed two polymorphic microsatellites in a candidate gene for fear response, monoamine oxidase A ( MAOA ), in chicken. Two hundred and eighty chickens from five breeds (Nagoya, Mikawa, White Leghorn, White Plymouth Rock, and Rhode Island Red) were investigated. A thymine (T) repeat and an adenine (A) repeat were found on MAOA intron 4 ( CMin4T ) and intron 9 ( CMin9A ), respectively. Nine alleles ( 127-bp to 166-bp ) in CMin4T and six alleles ( 184-bp to 198-bp ) in CMin9A were detected. A 128-bp allele in CMin4T was observed in the Nagoya breed only, implying a highly useful marker for discriminating the Nagoya breed from other breeds. In addition, the Mikawa breed had the fixed 127-bp and 198-bp alleles in CMin4T and CMin9A , respectively. The Nagoya breed chickens show densely cowardly behavior, but the Mikawa breed chickens do not show the same behavior. The current results may indicate that MAOA is an informative candidate gene for breed difference.     

Resistance phenotypes and genotypes among multiple-antimicrobial-resistant Salmonella enterica subspecies enterica serovar Choleraesuis strains isolated between 2008 and 2012 from slaughter pigs in Okinawa Prefecture,Japan

A total of 349 Salmonella enterica subspecies enterica serovar Choleraesuis (S. Choleraesuis) strains, which were isolated between 2008 and 2012 from 349 pigs at two slaughterhouses in Okinawa Prefecture, Japan, were investigated for antimicrobial susceptibility and the presence of antimicrobial resistance genes. All isolates were resistant to at least four antimicrobial agents. The antimicrobial agents for which isolates showed a high incidence of resistance were as follows: ampicillin (100%) and streptomycin (100%), followed by gentamicin (99.7%), oxytetracycline (99.7%), sulfamethoxazole/trimethoprim (99.4%), nalidixic acid (40.1%) and oxolinic acid (40.1%). All isolates were sensitive to cefuroxime, ceftiofur, colistin, fosfomycin, enrofloxacin, orbifloxacin and danofloxacin. The predominant resistance phenotypes and genotypes were: resistance to ampicillin, streptomycin, gentamicin, oxytetracycline and sulfamethoxazole/trimethoprim (58.5%, 204/349) and blaTEM-strA-strB-aadA1-aadA2-aacC2-tet (B)-sul1-sul2-dhfrXII-dhfrXIII (36.1%, 126/349). The quinolone resistance-determining regions (QRDRs) of gyrA, gyrB, parC and parE of the quinolone-resistant isolates (n=12) showed amino acid substitutions of Ser-83→Phe or Asp-87→Tyr in GyrA and Ser-107→Ala in ParC. To our knowledge, this is the first report on the molecular characterization of antimicrobial resistance among S. Choleraesuis strains in Japan.     

Cloning and expression analysis of prohibitin mRNA in canine mammary tumors

Prohibitin is an antiproliferative protein that is a product of a putative tumor suppressor gene. However, there is little information on prohibitins in companion animals. In this study, we cloned canine prohibitin mRNA using RT-PCR and 3′-RACE (Rapid Amplification of cDNA Ends). The sequence was well conserved compared with those of other mammals, including human. The deduced amino acid sequence translated from the open reading frame completely corresponded to the human sequence. Canine prohibitin mRNA was expressed in all normal mammary and tumor samples examined. These results suggest that this protein plays a vital role in cell growth mechanisms and may be related to the occurrence of canine mammary tumors.