Mitochondrial DNA analysis of Nepalese domestic dwarf cattle Lulu*

Dwarf Lulu cattle, the only Bos Taurus type of cattle in Nepal, are raised under severe environments in the mountainous zone of that country. In the present study, the body measurement traits, cytogenetic and molecular genetic characteristics of the Lulu cattle are investigated. Blood samples were collected from 31 animals in four villages (altitudes 2590–3550 m) in the southern part of Mustang. The Lulu cattle had a normal karyotype with 2n = 60, XY or XX. Only one male examined had a large submetacentric X‐chromosome and a small submetacentric taurine type Y‐chromosome. The mitochodrial DNA (mtDNA) genotypes were analyzed by PCR mediated restriction fragment length polymorphisms, displacement (D)‐loop region PCR mediated single strand conformation polymorphisms, and D‐loop region sequences. Many base substitutions were found in the D‐loop region, suggesting that the Lulu cattle originated from at least 10 maternal lines. Three types of mtDNA from these cattle were found, the Bos taurus type (n = 23), the Bos indicus type (n = 6), and the Bos grunniens type (n = 2). In the village at the lowest altitude, four of the five cows were of the Bos indicus type. These results indicated that mtDNA types of the Lulu cattle mostly belong to Bos taurus, but have been hybridized with Bos indicus cattle in lower‐elevation regions in their maternal lineage.     

Single oral β-cryptoxanthin administration increases its serum concentration and enhances peripheral blood neutrophil function in Holstein cattle

We investigated the effect of oral administration of β-cryptoxanthin (β-CRX) on its serum concentration and peripheral neutrophil functions by the chemiluminescence (CL) response in Holstein cattle. A single oral administration of β-CRX was performed for serum β-CRX concentration (0, 0.05, 0.1, or 0.2 mg/kg body weight [BW]) and for peak CL response of peripheral neutrophils (0.2 mg/kg BW). The serum β-CRX concentration was peaked on 2 days after, similar to peak CL response on 3 days after β-CRX administration. Therefore, a single oral administration of β-CRX (0.2 mg/kg BW) induces higher serum concentration and concurrently enhances bactericidal ability of peripheral neutrophils in Holstein cattle.     

Evaluation of truncated LipL32 expressed by Escherichia coli and Pichia pastoris for serodiagnosis of Leptospira infection in rodents

The applicability of the recombinant LipL32 for serodiagnosis of leptospiral infection in field rodents was assessed in this study. An immunodominant region of LipL32 was determined by monoclonal antibodies, and then, truncated LipL32 (tLipL32) was designed to contain the region (87–188th amino acid). The tLipL32 was compared between two recombinant expression hosts Escherichia coli and Pichia pastoris in ELISA. With field rat sera, tLipL32 expressed by P. pastoris (tLipL32p) had high antigenicity without background reactions, while tLipL32 expressed by E. coli (tLipL32e) showed high background reactions, which were reduced by pre-adsorption of sera with E. coli. To evaluate tLipL32-ELISA, field rat sera were tentatively divided into a Leptospira infection positive (12 sera) and a negative group (12 sera) based on the results from flaB gene PCR of kidney samples and WB with whole Leptospira cell. Consequently, the sensitivity of tLipL32p-ELISA for field rat sera was 83% . A similar result was obtained from tLipL32e-ELISA with adsorbed sera, (92%). However, sensitivity of tLipL32e-ELISA using sera without an adsorption treatment was 50%. Regardless of the expression host, tLipL32-ELISA had 100% specificity and sensitivity in experimentally infected laboratory rats. These results suggest that recombinant LipL32 expressed by P. pastoris is more applicable for serodiagnosis in field rats due to a lack of background reaction.     

Detection of CAP59 gene in 2 feline cases of systemic cryptococcosis

Two feline cases were diagnosed as systemic cryptococcosis due to Cryptococcus neoformans (teleomorph: Filobasidiella neoformans) by PCR assay with CAP59 gene primers using urine, serum and biopsy samples. The results of molecular analysis were consistent with the mycological findings.     

Uterine disorders diagnosed by ventrotomy in 47 rabbits

The type, ages of occurrence, primary complaints, clinical signs and mortality in forty-seven cases of uterine disorders diagnosed by ventrotomy in rabbits were analyzed retrospectively. Endometrial hyperplasia (29.8%) was most frequently observed, followed by uterine adenocarcinoma (21.3%). Tumorous lesions were seen in 46.8% of the cases. The age of occurrence ranged from two years and two months to seven years and six months, with a peak at four to five years of age. The most common primary complaint was bleeding (62.2%), followed by mammary gland abnormality (12.8%) and increased abdominal circumference (10.6%). Physical examinations revealed mammary gland disorders such as mammary cysts in 31.9% of the cases. Uterine disorders were detected by palpation in 15 out of 32 cases with a primary complaint of bleeding. Ultrasonography showed uterine disorders in 21 out of 24 cases, suggesting that ultrasonography could be useful in the diagnosis of uterine disorders. The outcome seemed to be influenced by physical status rather than malignancy of lesions. The mortality was higher in cases with symptoms such as anorexia, emaciation, severe anemia, and dehydration.     

Detoxification of α-tomatine by tomato pathogens Alternaria alternata tomato pathotype and Corynespora cassiicola and its role in infection

In tomato plants, α-tomatine, a steroidal glycoalkaloid saponin, inhibits fungal growth. Tomato pathogens that produce host-specific toxins, Alternaria alternata tomato pathotype causing Alternaria stem canker and Corynespora cassiicola causing Corynespora target spot, were investigated for sensitivity to α-tomatine. Although spore germination of A. alternata pathogenic and nonpathogenic to tomato and of C. cassiicola pathogenic to tomato was not affected by 0.1 mM α-tomatine, spore germination of C. cassiicola nonpathogenic to tomato was significantly inhibited. This result showed that A. alternata, regardless of its pathogenicity, and only the C. cassiicola pathogenic to tomato are resistant to α-tomatine. Germinating spores of A. alternata and C. cassiicola resistant to α-tomatine detoxified α-tomatine by degrading it to a less polar product. After inoculation of tomato leaves, spores of A. alternata and C. cassiicola nonpathogenic to tomato germinated and formed appressoria, but did not form infection hyphae in host tissues. When a host-specific toxin (CCT-toxin) produced by C. cassiicola pathogenic to tomato was added to nonpathogenic spores, colonization within leaves was observed in A. alternata, but not in C. cassiicola. On the other hand, when spores of C. cassiicola nonpathogenic to tomato were suspended in spore germination fluid of nonpathogenic A. alternata with α-tomatine detoxification activity, the fungus could be induced to colonize leaves in the presence of CCT-toxin. These results indicate that A. alternata tomato pathotype and C. cassiicola pathogenic to tomato detoxify α-tomatine during infection and that this detoxification is essential for host colonization by pathogens that produce host-specific toxins.     

Influence of feeding regime on timing of parturition in beef cattle and the relationship of vaginal temperature to parturition

The timing of parturition was recorded for a total of 56 beef cattle (Japanese Black × Holstein Friesian) on different dietary treatments. The rate of calving during daylight hours in cows night‐fed (18.00 hours) with a roughage diet was significantly higher than that in cows night‐fed with a high concentrate diet (79.2% vs 38.5%, P < 0.05). Subsequently, the vaginal temperature (VT) of these cows was analyzed using a cosinor method. When the feeding schedule was changed from twice daily (08.30 and 15.30 hours) to night feeding, the periodicity, the acrophase and the bathyphase, which were the parameters of the cosine curve, were unstable from the first day of night feeding until after day 6 (P < 0.05). Prior to parturition, the midline‐estimating statistic of rhythm (MESOR) and the amplitude for the cows that were fed a high‐roughage diet at night and that calved at night‐time were lower and larger, respectively, than that for the other treatments (P < 0.01). Based on these results, the time of parturition in most of the beef cows was influenced by feeding time and diet composition. Those cows that calved at night‐time in spite of night feeding had lower vaginal temperatures.     

The effect of topical minoxidil treatment on hair follicular growth of neonatal hairless descendants of Mexican hairless dogs

The effect of daily topical minoxidil treatment on hair growth was investigated in eight neonatal hairless pups. After 21 days of minoxidil treatment, hair growth occurred in the minoxidil-treated areas of each neonatal hairless pup. The numbers of hairs in the treated areas increased as compared with that in the areas treated with vehicle only. Histological findings revealed that the skin treated with minoxidil contained many hair follicles derived from epidermal ingrowths (hair germs) projecting into the dermis. These hair follicles showed various stages of neofolliculogenesis. After 31 days of treatment, terminal hair growth was observed. In the minoxidil-untreated areas, epidermal ingrowths remained undifferentiated. These results revealed that hairless dogs developed hair growth reactions following minoxidil-treatment when such treatment is applied in the early neonatal period. Neonatal hairless pups are a useful model for evaluating the effectiveness of hair growth stimulators.