Use of quantitative real-time RT-PCR to investigate the correlation between viremia and viral shedding of canine distemper virus,and infection outcomes in experimentally infected dogs

We used real-time RT-PCR and virus titration to examine canine distemper virus (CDV) kinetics in peripheral blood and rectal and nasal secretions from 12 experimentally infected dogs. Real-time RT-PCR proved extremely sensitive, and the correlation between the two methods for rectal and nasal (r=0.78, 0.80) samples on the peak day of viral RNA was good. Although the dogs showed diverse symptoms, viral RNA kinetics were similar; the peak of viral RNA in the symptomatic dogs was consistent with the onset of symptoms. These results indicate that real-time RT-PCR is sufficiently sensitive to monitor CDV replication in experimentally infected dogs regardless of the degree of clinical manifestation and suggest that the peak of viral RNA reflects active CDV replication.     

Identification and expression of a gene encoding an epitope that induces hemagglutination inhibition antibody to Avibacterium paragallinarum serovar A

The aims of this study were the identification, cloning, and expression of a genetic region encoding an epitope that induces hemagglutination inhibition (HI) antibody against Avibacterium paragallinarum serovar A and an evaluation of the recombinant protein for immunogenicity in chickens. Although two monoclonal antibodies (MAbs) with HI activity, designated S24-951 and S7-1716-5C, were generated in this study, no reactive proteins with both MAbs were identified by Western blot analysis. A gene fragment of 5157 bp, designated hpa5. 1, was cloned from genomic DNA, and a recombinant protein expressed by hpa5.1, designated HPA5.1, reacted with both MAbs on dot-blot analysis. HPA5.1 showed no hemagglutinating activity, but significantly absorbed HI antibodies in the chicken immune serum. Analysis using a series of deletion mutants prepared from hpa5.1 indicated that a 4.8 kbp gene in hpa5.1 is essential for the expression epitope recognized by MAb S24-951. In addition, chickens immunized once with HPA5.1 showed a high protection rate with sufficient HI antibody titers against challenge exposure with a virulent strain of A. paragallinarum serovar A strain 221. These results show that hpa5. I1 is responsible for the expression of an epitope that induces HI antibody, and HPA5.1 might be a candidate for the development of a new vaccine against avian infectious coryza caused by A. paragallinarum serovar A.     

Breeding season in female cats acclimated under a natural photoperiod and interval until puberty

The breeding season was investigated in 174 female cats that were acclimated under a natural photoperiod, and determined the interval between birth and initial estrus (puberty) was determined in 125 cats. Although the breeding season differed noticeably among individual animals, the mean was 180.4 +/- 3.0 (SE) days between the end of January and the end of July. The interval between birth and first estrus ranged from 181 to 560 days, with a mean of 345.0 +/- 0.9 days. With respect to month of birth, the mean interval was 343.0 +/- 9.5 days in cats born between March and June. Among cats that were born between July and October, the mean intervals were 242.0 +/- 6.3 days in cats that exhibited estrus the year after birth and 519.2 +/- 5.8 days in those that exhibited estrus 2 years after birth.     

Disappearance of the PHA-E lectin binding site on the surface of ejaculated sperm and sperm capacitation in the dog

Ejaculated semen and cross sections of the cauda epididymides collected from 3 normal dogs were smeared or stamped on glass slides, and the sperm on the slides were stained with 7 different FITC-lectins (Con A, DBA, GS-1, PHA-E, PSA, UEA-1, WGA) to examine the relation between sperm-binding glycoprotein derived from the canine prostate and sperm capacitation. The only lectin that stained the ejaculated sperm but not the cauda epididymal sperm was PHA-E. The sperm ejaculated by 5 other dogs were incubated for 4 hr in fluid flushed from the uterine horns or oviducts of estrous bitches, and then the percentages of actively motile sperm and hyperactivated sperm (HA-sperm) were determined. The percentages of PHA-E-labeled sperm and sperm labeled with fluoresceinated Ca indicator to assess the influx of Ca into the sperm were also evaluated. The mean percentages of actively motile sperm, HA-sperm, and Ca-labeled sperm after 4 hr of incubation in the uterine flush fluid and oviductal flush fluid were significantly higher than in control medium (P<0.05, 0.01), but the mean percentages of PHA-E-labeled sperm were lower (both P<0.01). The percentages of PHA-E-unlabeled sperm correlated with the percentages of both HA-sperm and Ca-labeled sperm (r(2)=0.787 and 0.812, respectively). The results indicate that loss of the glycoprotein secreted by the canine prostate on the sperm surface induces the influx of Ca into the sperm, and then hyperactivation of the sperm.     

Formation of detached tail and coiled tail of sperm in a beagle dog

The right testis and epididymis were excised from a Beagle dog that ejaculated high percentages of sperm with detached tails and with coiled tails. Cross sections of the organs were stamped on glass slides and histological examination of the organs was performed to find the portion where sperm with the abnormal tails appear. Many sperm with tails whose axoneme was exposed near the neck region were observed in the testis and they decreased in order from the caput, to the corpus, and the cauda epididymis. Sperm with detached tails and sperm with coiled tails gradually increased in the epididymis. These findings indicate that the tails of sperm with an exposed axoneme detached in the epididymis.     

Changes in plasma testosterone levels and semen quality after 3 injections of a GnRH analogue in 3 dogs with spermatogenic dysfunction

Three male Beagles with low plasma luteinizing hormone and testosterone levels and spermatogenic dysfunction (SD-dogs) were given 3 weekly subcutaneous injections of a gonadotropin-releasing hormone analogue (Buserelin; GnRH-AB). The plasma T level of all of the SD-dogs increased and peaked at 4-6 weeks after the first injection. The total number of sperm and the sperm motility of the three SD-dogs increased 5-7 weeks after the first injection. Therefore, the authors concluded that 3 weekly injections of GnRH-AB transiently improves semen quality in some cases of canine spermatogenic dysfunction.     

Genetic homogeneity between adult and juvenile populations of <Emphasis Type="Italic">Scombrops gilberti</Emphasis> (Percoid,Scombropidae) in the Pacific Ocean off the Japanese Islands

Scombrops gilberti is a member of the percoid family Scombropidae, which includes a single genus and three to four species worldwide. Little is known about the ecology of this species. Juvenile S. gilberti have been found in the waters off northern Japan (Iwate Prefecture), whereas adults are found in the waters around the southern counterpart (Izu Islands), approximately 700 km from the northern waters. In the present study, we captured immature S. gilberti (106–248 mm standard length, SL) in the northern waters by set net at 8–80 m depth, whereas larger individuals (150–328 mm SL) were captured by trawling at 150–500 m depth. By contrast, only adult S. gilberti (422–590 mm SL) were captured in the southern waters. The genetic composition of the adult population of S. gilberti from the southern waters and of the juvenile population from the northern waters was compared using the nucleotide sequence of the mitochondrial DNA (mtDNA) cytochrome b gene. No significant differences in genetic parameters such as fixation index, neutrality test or mismatch distribution analysis were found between these geographically distinct populations of S. gilberti, showing that these populations are genetically homogeneous.