Transmissible gastroenteritis in neonatal dogs: experimental intestinal infection with transmissible gastroenteritis virus.

Fourteen neonatal dogs (4 through 11 days of age) were exposed orally to the Purdue strain of transmissible gastroenteritis (TGE) virus, and six dogs of similar age were noninoculated controls. Clinical signs of enteric disease did not develop. Both exposed and control dogs had normal fecal passages and appetite throughout the experiment. Jejunal epithelium from dogs euthanatized at 12, 24, 48, and 96 hours and at 10 days after exposure did not exhibit morphologic alterations detectable by light microscopy. Electron microscopic examination indicated that jejunal epithelial cells contained TGE viral particles as early as 12 hours after dogs were exposed. There were no apparent morphologic alterations or signs of desquamation of virus-infected cells, however. Results of pig transmission studies indicated that viable TGE virus was in jejunal tissue of the dogs as early as 12 hours and as late as 10 days after exposure to the virus.     

Polyarteritis in a beagle

A 13-month-old Beagle became anorectic and had fever, stiff gait, and tenderness in the inguinal region. Clinical signs of disease were associated with neutrophilia and a decrease in the albumin-to-globulin ratio. The dog became clinically normal for 5 days after 3 days of treatment with penicillin G and dihydrostreptomycin. Clinical signs of disease recurred, and the dog was euthanatized after failing to respond to administration of a trimethoprim-sulfamethoxazole combination for 9 days. Disseminated arteritis was seen in the testes, epididymides, mesentery, coronary arteries, aorta, and thyroid gland. Lesions were seen in large and medium-sized arteries and varied from acute necrotizing arteries to a chronic lesion with organization and recanalization of thrombi. The clinical signs of disease resembled those of Beagle pain syndrome, described in laboratory Beagles.     

Simultaneous temporal progress of sorghum anthracnose and leaf blight in crop mixtures with disparate patterns

Field studies were conducted at Alupe in western Kenya in 1995 and 1996 to evaluate the efficacy of crop and species mixtures for the management of sorghum anthracnose (caused by Colletotrichum sublineolum) and leaf blight (caused by Exserohilum turcicum). The progress of these diseases developing simultaneously on a susceptible sorghum cultivar planted in inter- or intra-row mixtures of varying proportions with either maize or resistant sorghum was monitored. The effects of host type and mixture patterns on disease progress were compared by parameter estimates derived from fitted Lotka-Volterra competition equations and nonlinear logistic models. Competition coefficients were not significant and their confidence intervals included zero in most cases, suggesting that interactions between C. sublineolum and E. turcicum did not occur. Mixtures of the susceptible sorghum with either the nonhost maize or the resistant sorghum delayed the time when disease is first observed and reduced the rate of disease progress and carrying capacity for both anthracnose and leaf blight, with a more pronounced effect on the latter disease. The lower efficacy of mixtures in reducing anthracnose was attributed to an aggregated spatial pattern, coupled with higher rates of progress for this disease. Intra-row mixtures were more efficient than inter-row mixtures in reducing disease development in all years. The implications of these observations for the management of sorghum diseases under small-scale farming systems are discussed.     

Regulation of urease production in soil by microbial assimilation of nitrogen

Summary Studies of the effects of different forms of N on urease production in soils amended with organic C showed that although microbial activity, as measured by CO₂ production, was stimulated by the addition of NH₄ ⁺ or NO₃ ^- to C-amended soils (200 mol glucose-C g^–1 soil), urease production was repressed by these forms of N. The addition of L-methionine sulfoximine, an inhibitor of inorganic N assimilation by microorganisms, relieved the NH₄ ⁺ and NO₃ ^- repression of urease production in C-amended soil. The addition of sodium chlorate, an inhibitor of NO₃ ^- reduction to NH₄ ⁺ by microorganisms, relieved the NO₃ ^- repression of urease production, but did not eliminate the repression associated with NH₄ ⁺. These observations indicate that microbial production of urease in C-amended soils is not directly repressed by NH₄ ⁺ or NO₃ ^-, but by products formed by microbial assimilation of these forms of N. This conclusion is supported by our finding that the biologically active L-isomers of alanine, arginine, asparagine, aspartate, and glutamine, repressed urease production in C-amended soil, whereas the D-isomers of these amino acids had little or no influence on urease production. This work suggests that urease synthesis by soil microorganisms is controlled by the global N regulon.     

Safety of an attenuated West Nile virus vaccine, live Flavivirus chimera in horses

REASON FOR PERFORMING STUDY: West Nile virus (WNV) infection is endemic and able to cause disease in naive hosts. It is necessary therefore to evaluate the safety of new vaccines. OBJECTIVES: To establish: 1) the safety of a modified live Flavivirus/West Nile virus (WN-FV) chimera by administration of an overdose and testing for shed of vaccine virus and spread to uninoculated sentinel horses; 2) that this vaccine did not become pathogenic once passaged in horses; and 3) vaccine safety under field conditions. METHODS: There were 3 protocols: 1) In the overdose/shed and spread study, horses were vaccinated with a 100x immunogenicity overdose of WN-FV chimera vaccine and housed with sentinel horses. 2) A reversion to virulence study, where horses were vaccinated with a 20x immunogenicity overdose of WN-FV chimera vaccine. Horses in both studies were evaluated for abnormal health conditions and samples obtained to detect virus, seroconversion and dissemination into tissues. 3) In a field safety test 919 healthy horses of various ages, breeds and sex were used. RESULTS: Vaccination did not result in site or systemic reactions in either experimental or field-injected horses. There was no shed of vaccine virus, no detection of vaccine virus into tissue and no reversion to virulence with passage. CONCLUSIONS: WN-FV chimera vaccine is safe to use in horses with no evidence of ill effects from very high doses of vaccine. There was no evidence of reversion to virulence. In addition, administration of this vaccine to several hundred horses that may have been previously exposed to WNV or WNV vaccine resulted in no untoward reactions. POTENTIAL RELEVANCE: These studies establish that this live attenuated Flavivirus chimera is safe to use for immunoprophylaxis against WNV disease in horses.     

Randomly primed,strand-switching,MinION-based sequencing for the detection and characterization of cultured RNA viruses

RNA viruses rapidly mutate, which can result in increased virulence, increased escape from vaccine protection, and false-negative detection results. Targeted detection methods have a limited ability to detect unknown viruses and often provide insufficient data to detect coinfections or identify antigenic variants. Random, deep sequencing is a method that can more fully detect and characterize RNA viruses and is often coupled with molecular techniques or culture methods for viral enrichment. We tested viral culture coupled with third-generation sequencing for the ability to detect and characterize RNA viruses. Cultures of bovine viral diarrhea virus, canine distemper virus (CDV), epizootic hemorrhagic disease virus, infectious bronchitis virus, 2 influenza A viruses, and porcine respiratory and reproductive syndrome virus were sequenced on the MinION platform using a random, reverse primer in a strand-switching reaction, coupled with PCR-based barcoding. Reads were taxonomically classified and used for reference-based sequence building using a stock personal computer. This method accurately detected and identified complete coding sequence genomes with a minimum of 20× coverage depth for all 7 viruses, including a sample containing 2 viruses. Each lineage-typing region had at least 26× coverage depth for all viruses. Furthermore, analyzing the CDV sample through a pipeline devoid of CDV reference sequences modeled the ability of this protocol to detect unknown viruses. Our results show the ability of this technique to detect and characterize dsRNA, negative- and positive-sense ssRNA, and nonsegmented and segmented RNA viruses.     

Outcomes after extrahepatic portosystemic shunt ligation in 49 dogs

OBJECTIVE: To evaluate outcomes after attenuation of extrahepatic portosystemic shunts in dogs using surgical silk. DESIGN: Retrospective study. PROCEDURE: Case records were reviewed for degree of surgical attenuation, experience of the primary surgeon, perioperative mortality and problems related to persistent portosystemic shunting or shunt ligation. Presence of portosystemic shunting after surgery was evaluated by ammonia tolerance testing, measurement of postprandial serum bile acid, plasma urea and cholesterol concentrations and liver enzyme activity. The influence of age, postocclusion portal pressure, primary surgeon, degree of attenuation and postoperative biochemical findings on the occurrence of postoperative problems was assessed. RESULTS: The mortality rate was 2.1%. Shunt attenuation was complete in 34% and partial in 66% of dogs. Portal hypertension necessitating ligature removal was encountered in only one dog. Five dogs experienced neurological abnormalities (seizures or ataxia), possibly as a manifestation of 'postligation seizure syndrome'. Postoperative liver function was normal in 78% of dogs, including 70% with partial shunt attenuation. Experience of the surgeon was related positively to outcome after partial attenuation (P = 0.002). Postoperative biochemical evidence of abnormal liver function was the most sensitive predictor of recurrence of clinical signs referable to persistent portosystemic shunting. CONCLUSIONS: In the hands of an experienced surgeon, surgical attenuation of single extrahepatic shunts was safe and effective, even in animals with partial attenuation. Most dogs with biochemical evidence of persistent shunting suffer relapse of clinical signs within 18 months of surgery. Postligation neurological syndromes of variable intensity may be more common than previously thought.     

Effect of oxytocin on expression of cytosolic phospholipase A2 mRNA and protein in ovine endometrial tissue in vivo

The induction of endometrial prostaglandin (PG) F₂ synthesis by oxytocin is dependent upon activation of phospholipase (PL) A₂ and mobilization of arachidonic acid. The objective of this study was to determine if oxytocin stimulates PGF₂ synthesis by inducing synthesis of cytosolic PLA₂ (cPLA₂). In Experiment 1, 15 ovariectomized ewes were given progesterone and estradiol to simulate an estrous cycle. Ewes were then given an injection of oxytocin on Day 14 of the simulated estrous cycle. Jugular blood samples were collected and assayed for 13,14-dihydro-15-keto-prostaglandin F₂ (PGFM). Uteri were collected at 0, 7.5, 25, 90, or 240 min postinjection (n = 3 ewes/time point). Total RNA was isolated from caruncular endometrium and subjected to dot-blot analysis. Oxytocin induced a rapid and transient increase in serum PGFM (P < 0.01). However, endometrial concentrations of cPLA2 mRNA did not change following oxytocin administration (P > 0.10). In Experiment 2, 11 ovary-intact ewes were given oxytocin (n = 5) or saline (n = 6) on Day 15 after estrus. Jugular blood samples were collected and assayed for serum concentrations of PGFM. Uteri were collected at 15 min postinjection. Homogenates were prepared from caruncular endometrium and subjected to Western blot analysis. Concentrations of PGFM were higher in oxytocin treated ewes compared to saline treated ewes at 15 min postinjection (P < 0.01). Endometrial concentrations of cPLA₂ protein were greater in the cytosolic than in the microsomal fraction (P < 0.01). Oxytocin did not affect the amount of cPLA₂ protein in either fraction (P > 0.10). In conclusion, oxytocin did not effect expression of either cPLA₂ mRNA or protein in ovine endometrium. Oxytocin may stimulate PGF₂ synthesis by activating cPLA₂ protein that is already present in an inactive form.     

Technical note: Constrained optimization of breed composition in composite populations to balance net merit and risk

We developed a method to derive optimum composite genotypes when the proportion of a specific breed (or combination of breeds) in the composite is constrained to a desired level. The method allows development of composites that balance net merit according to available parameters and reflects the production system and(or) risk preference, particularly when traits in the breeding objective do not represent all traits important for production. The method can also be used to evaluate the cost of constraining breed use. For example, when the true optimal composite is difficult to obtain in the time frame available to the producer, the predicted merit of the composite with breed proportion restricted to those that are obtainable in practice can be compared with the predicted merit of the true optimum composite. In a case study the method is used to optimize a tropical composite using Brahman, Charolais, and Belmont Red breeds. The proportion of Charolais is to be constrained to 50%. Using growth parameters from a tropical crossbreeding experiment, the unconstrained optimum composite was 32.5% Brahman, 11.1% Belmont Red, and 56.3% Charolais. When the proportion of Charolais in the composite was constrained to 50%, optimum proportions of Brahman and Belmont Red were 33.9% and 16.1% respectively. In the case study net merit (annualized profit per cow exposed, $), as defined by the growth parameters, was .2% less than that of the unconstrained optimum composite.