Molecular cloning of canine thrombomodulin cDNA and expression in normal tissues

Thrombomodulin (TM) is a glycoprotein localized mainly on endothelial cell surfaces, and is a major regulator of vascular thromboresistance. The entire open reading frame of canine TM cDNA comprises 1737 bp, encoding 578 amino acid residues. Comparison of the deduced amino acid sequence from canine TM with those of human, mouse, rat, rabbit and bovine (partial) TM sequences revealed 73.1%, 69.1%, 65.8%, 74.3% and 69.5% identity, respectively. Canine TM mRNA expression was confirmed by RT-PCR analysis in lung, liver, spleen, kidney, pancreas and lymph node, and was relatively low in heart, cerebrum, urinary bladder and uterus. The present results provide valuable data for research into canine coagulation disorders.     

Serological differentiation of FIV-infected cats from dual-subtype feline immunodeficiency virus vaccine (Fel-O-Vax FIV) inoculated cats

Feline immunodeficiency virus (FIV) vaccine, Fel-O-Vax FIV, was released for sale in the US in 2002. The antibodies of vaccinated cats interfere with serological assays by currently available FIV diagnostic kits. In this study, we investigated whether it is possible to distinguish serologically cats vaccinated with Fel-O-Vax FIV from cats experimentally or naturally infected with FIV. A total of 153 sera taken from 97 cats were used as serum samples. Enzyme linked immunosorbent assay (ELISA) was performed using whole FIV antigen and formalin treated whole FIV antigen, recombinant-gag (r-gag) antigen, and transmembrane (TM) peptide. Statistical analysis was performed using ELISA optical density (O.D.) values obtained with each antigen as variables. Except for the ELISA O.D. values obtained with r-gag antigen, a significant difference in ELISA O.D. values was observed between the vaccinated and the infected groups. However, it was not possible to distinguish both groups unequivocally. Using discriminant analysis, it was possible to distinguish the two groups with an accuracy of 97.1% with two discriminating variables (ELISA O.D. values obtained with formalin treated whole FIV antigen, and TM peptide), 97.8% with three discriminating variables (ELISA O.D. values obtained with whole FIV antigen, formalin treated whole FIV antigen, and TM peptide). Therefore, it was considered possible to distinguish cats vaccinated with Fel-O-Vax FIV from FIV-infected cats by ELISA using two types of antigens including formalin treated whole FIV antigen and TM peptide, or three types of antigens including formalin treated whole FIV antigen, TM peptide and whole FIV antigen.     

Operation of the Ba Lai irrigation system in the Mekong Delta, Vietnam

Ba Lai Irrigation Project is located at coastal area in the Mekong Delta. In dry season when the flow rate decreases and the strong east wind blows into the delta, the salinity intrusion increases and seriously affects agricultural and domestic water use. Intakes of Ba Lai system have to be closed for 1–3 months depending on their locations, and no water supply during this period often causes water pollution in the project area. In order to solve such problems, this study aims to seek gate operation procedures for salinity control and water environment improvement. A numerical model is developed to simulate water movement, salinity concentration and duration of remaining water (water age) within the system under three scenarios: (1) without control structures, (2) with available control structures, (3) with the full control structures. Through the numerical simulations, control structures are confirmed to be an effective measure for the salinity control and suitable gate operation schedules are proposed to improve the water environment in the project.     

Induction of chemoresistance in a cultured canine cell line by retroviral transduction of the canine multidrug resistance 1 gene

OBJECTIVE: To induce chemoresistance in a normal canine cell line through the transduction of the canine multidrug resistance 1 gene (mdr1). SAMPLE POPULATION: Madin-Darby canine kidney (MDCK) epithelial cell line. PROCEDURES: The full-length canine mdr1 cDNA clone isolated in our laboratory was inserted into a Moloney murine leukemia virus-based vector to construct the retroviral vector, pLNC-cMDR1. After retroviral transduction of pLNC-cMDR1 into MDCK cells, the expression and function of the P-glycoprotein, a product of mdr1, were assessed by immunoblotting, measurement of rhodamine123 (Rh123) retention, and drug sensitivity assays. RESULTS: P-glycoprotein was strongly expressed in cells transduced with pLNC-cMDR1. This P-glycoprotein was fully functional, as demonstrated by the decreased Rh123 retention and the increased resistance to chemotherapeutic drugs. Measured as 50% inhibitory concentrations, resistance increased 59 times to vincristine and 25 times to doxorubicin in MDCK cells after transduction of pLNC-cMDR1. CONCLUSIONS AND CLINICAL RELEVANCE: Transduction of canine mdr1 is an effective method for inducing chemoresistance in normal canine cells. This system may be applicable to the induction of drug resistance in hematopoietic cells.     

Comparative study on yield and chemical composition of maize (Zea mays L.) and sorghum (Sorghum bicolor Moench) using different levels of manure application

The effects of three levels of manure application (0, 4 and 8 ton/10 a) with basic fertilizer (N : P : K = 10:10:10 kg/10 a) on the yield and chemical composition of maize (Pioneer 32K61) and sorghum (FS501) forages were examined in 2 successive years, 2001–2002. Maize was harvested at yellow ripe stage, while sorghum at dough stage. Dry matter yield (DMY) of sorghum was significantly higher than that of maize in both 2001 and 2002. Although, DMY of both sorghum and maize were not significantly affected by manure levels in 2001, they increased with the level of manure in 2002. Maize had higher levels of crude protein (CP) and lower crude fiber concentration than those of sorghum in both 2001 and 2002. Manure application had no effect on chemical compositions of both maize and sorghum except for CP in both years and organic a fraction (Oa) concentration in 2002. The higher manure application might have greater effects on sorghum and maize production because of higher DMY in the second year without any increase in fiber fraction.     

Ebstein's anomaly in a beagle dog

An intact male beagle dog aged 1 year was referred because of shortness of breath, exercise intolerance and cardiac murmur. Based on the results from electrocardiography, thoracic radiography and echocardiography, the dog was diagnosed as Ebstein's anomaly. Although the orally administered digoxin, vasodilators and diuretics partially improved congestive signs, the dog became to be refractory and died 20 months after the diagnosis. Necropsy confirmed malformation and apical displacement of the basal attachment of tricuspid valve leaflets.     

Antagonistic effects of atipamezole and yohimbine on medetomidine-induced diuresis in healthy dogs

This study aimed to investigate and compare the antagonistic effects of atipamezole and yohimbine on medetomidine-induced diuresis in healthy dogs. Five dogs were used repeatedly in each of 8 groups. One group was not medicated. Dogs in the other groups received 20 μg/kg of medetomidine intramuscularly and, 0.5 h later, saline (as the control injection), 50, 100, or 300 μg/kg of atipamezole, or 50, 100, or 300 μg/kg of yohimbine intramuscularly. Urine and blood samples were taken 11 times over 24 h for measurement of the following: urine volume, specific gravity, and creatinine concentration; urine and plasma osmolality; urine and plasma concentrations of electrolytes and arginine vasopressin (AVP); and the plasma concentration of atrial natriuretic peptide (ANP). Both atipamezole and yohimbine antagonized the diuretic effect of medetomidine, inhibiting medetomidine-induced decreases in urine specific gravity, osmolality, and concentrations of creatinine, sodium, potassium, chloride, and AVP and reversing both the medetomidine-induced increase in plasma concentrations of sodium, potassium, and chloride and the medetomidine-induced decrease in the plasma AVP concentration. Atipamezole significantly stimulated ANP release. The antidiuretic action of yohimbine was more potent than that of atipamezole but was not dose-dependent, in contrast to the action of atipamezole. The effects of these drugs may not be due only to actions mediated by α₂-adrenoceptors.     

Prevalence and risk factors associated with Cryptosporidium oocysts shedding in pigs in Central Vietnam

We investigated the prevalence and risk factors associated with Cryptosporidium oocysts shedding in pigs in Central Vietnam. A total of 740 single fecal samples collected from diarrheic and non-diarrheic pigs on 89 farms were screened by the modified Ziehl-Neelsen staining method. Prevalence at the animal and the farm levels were 18.1% (134/740) and 71.9% (64/89), respectively. Risk factors for the infection were identified using univariate and multivariate logistic regression analysis. The results revealed that age, sanitary condition and topography were significantly associated with oocyst shedding (P<0.05). Pre-weaned piglets were at the highest risk for infection, followed by post-weaners, sows and finishing pigs. Good sanitary conditions showed positive effects in decreasing oocysts shedding. Topographically, Cryptosporidium was more common in mountainous zone than that in coastal delta zone. There was an association between the occurrence of diarrhea and the level of Cryptosporidium oocyst excretion within infected pigs. This is the first epidemiological investigation of prevalence and risk factors of Cryptosporidium in pigs in Vietnam.     

The formation process of button ulcers in pigs experimentally infected with a subgenotype 2.1 isolate of classical swine fever virus

We evaluated the role of classical swine fever virus (CSFV) in the formation of button ulcers in the mucosa of the gastrointestinal tract. Histopathological and immunohistochemical analyses of pigs experimentally infected with a subgenotype 2.1 isolate of CSFV, which was isolated in Japan in 2019, revealed follicular necrosis in the submucosal mucosa-associated lymphoid tissue and herniation of crypts as factors that contribute to the development of button ulcers during CSFV infection. These findings indicate that CSFV induces follicular necrosis and is one of the causative agents of button ulcers in pigs.