Serum complement activity in the three-toed amphiuma (Amphiuma tridactylum)

Some animals routinely endure serious injuries from predators or during intraspecific territorial conflicts. Such is the case for Amphiuma tridactylum, an aquatic salamander that lives in an environment rich in potentially infectious microbes, apparently with rare or no pathogenic infection. Some vertebrates possess innate immune mechanisms, but whether this is the case for Amphiuma is unknown. To assess this potential, plasma from 19 A. tridactylum was pooled and used for characterisation of serum complement activity. The ability of A. tridactylum plasma to hemolyse unsensitised sheep red blood cells (SRBCs) was titer-dependent, with low activity observed even at high plasma titers. The kinetic characterisation of SRBC hemolysis revealed that significant activity could be measured within 10 min of incubation, and maximal activity occurred within 60 min. The SRBC hemolysis by A. tridactylum plasma was also temperature-dependent, with maximal activity at 30 °C. In addition, this activity was sensitive to mild heat treatment, with 96% of activity inhibited by incubation at 56 °C for 30 min. The SRBC hemolysis could also be inactivated by pretreatment of the plasma with proteases, indicating that this activity was protein dependent. The activity required divalent metals ions, with activity inhibited by EDTA, citrate, or phosphate. However, the chelator-inhibited activity could be restored by the addition of excess Ca²⁺ or Mg²⁺, but not Cu²⁺ or Ba²⁺, indicating specificity of the divalent metal ion requirement. The sensitivity to heat, proteases, and divalent metal ion chelators strongly suggests that A. tridactylum plasma-mediated hemolysis of SRBCs is mediated by the serum complement system of proteins.     

Resynchronization of estrus in beef cattle: ovarian function, estrus and fertility following progestin treatment and treatments to synchronize ovarian follicular development and estrus

The objective was to optimize rebreeding of nonpregnant, previously inseminated beef cattle. In Experiment 1, 43 cows received a used intravaginal progesterone-releasing insert (IVPRI; Days 0-7) 12.3 d after ovulation and received concurrently no treatment, 100 microg gonadotropin releasing hormone (GnRH), 1 mg estradiol cypionate (ECP), or 150 mg progesterone. Emergence of a new ovarian follicular wave was most synchronous (P < 0.0001) in the GnRH group. In Experiment 2, 675 heifers were given GnRH or no treatment on Day 0, fed melengestrol acetate (MGA; 0.5 mg/head/d) from Days 0-5 (Day 0 = 13-14 d after timed insemination; TAI), given 0.5 mg ECP or nothing on Day 7, and reinseminated 6-12 h after onset of estrus. Estrus was more synchronous (P < 0.05) in heifers given GnRH versus no treatment on Day 0. In Experiment 3, 317 TAI heifers were resynchronized with either MGA or a used IVPRI with or without ECP on Day 7; estrus was more synchronous (P < 0.05) and pregnancy rates were higher (54.1% versus 39.2%, P < 0.05) in heifers given a used IVPRI than those fed MGA. For resynchronization of heifers, pregnancy rates were not significantly improved with GnRH treatment, but were higher with a used IVPRI than with MGA.     

A multivariable prognostic model for equine colic patients

A survey of 1965 equine colic cases was conducted from August 1985 to July 1986 ten equine referral hospitals located througout the U.S.A. Two-thirds of the cases were randomly selected for model development (1336), while the remaining cases (629) were used only for subsequent validation of the model. The following outcomes were defined: (1) died or killed prior to discharge from the hospital; or (2) alive at the time of discharge. Only variables which were significant (P<0.05) in an initial bivariate screening procedure and for which there were less than 400 missing values were considered in the multivariable modelling. A multivariable logistic regression model was constructed using a stepwise algorithm. The model used 666 cases and included the following variables: peripheral pulse (normal or weak), pulse rate, surgical or medical treatment, packed cell volume, self-inflicted trauma (absent or present) and capillary refill time. For each horse in the validation data set which had the appropriate variables recorded (n=335), the estimated expected probability of death (expected value) was calculated using the logistic regression equation. Using Bayes theorem, the post-test probability was calculated from the expected value (an estimate of the test odds) and the present probability (the case-fatality rate at each institution). Nomograms of predictive values for different case-fatality rates and expected values were constructed. Hosmer-Lemeshow goodness-of-fit statistics indicated that the model fitted the model data set well but the validation set poorly. However, when the observed case-fatality rates were compared with the average post-test probabilities for 0.10 increments of post-test probability, qualitatively, the model's performance was better.     

Generation of Salmonella ghost cells expressing fimbrial antigens of enterotoxigenic Escherichia coli and evaluation of their antigenicity in a murine model

Salmonella Typhimurium ghost cells expressing K88ab, K88ac, K99, and FasA fimbriae of enterotoxigenic Escherichia coli (ETEC) in their envelopes were constructed. The genes encoding the fimbriae were individually cloned into an expression plasmid, pMMP81, carrying the asd gene, which was subsequently electroporated into the Δasd S. Typhimurium mutant. Plasmid pJHLP99, carrying the phiX174 lysis gene E, was also subsequently electroporated into the Salmonella mutant. The presence of the individual fimbriae on the ghost cells was examined by Western blot analysis. Forty BALB/c mice were equally divided into 2 groups of 20 mice each. Group A mice were intramuscularly vaccinated with a mixture of the 4 ghost cells expressing the individual fimbriae. The group B mice were inoculated with sterile phosphate-buffered saline as a control. The antigen-specific serum IgG concentrations were significantly higher in group A than in group B from week 2 until week 6 after inoculation. In addition, the antigen-specific IgA concentrations in fecal samples were significantly higher in group A than in group B at week 2 after inoculation. A large difference between the groups in the number of antigen-specific IgA-secreting cells in the small intestine was observed by immunohistochemical study. Also, the splenic lymphocyte proliferative responses were significantly greater in group A than in the control mice. These results suggest that vaccination with our Salmonella ghost cells can induce both humoral and cell-mediated immune responses and that the increased number of antigen-specific IgA-secreting cells in the small intestine may be correlated with the elevated fecal IgA immune response.     

Evaluation of an air-filtration system for preventing aerosol transmission of Porcine reproductive and respiratory syndrome virus

The purpose of this study was to evaluate the ability of a commercial air-filtration system to reduce aerosol transmission of Porcine reproductive and respiratory syndrome virus (PRRSV). The system consisted of a pre-filter and 2 filters with EU8 and EU13 ratings. In each of 4 trials, 5 PRRSV-infected donor pigs and 1 naive recipient pig (each 25 kg) were housed in opposing chambers connected by a 1.3-m-long duct. The system filtered air entering 1 recipient-pig chamber (filtered facility) from the donor-pig chamber but not a 2nd recipient-pig chamber (nonfiltered facility). The donor pigs had been experimentally infected with PRRSV MN-184, an isolate previously documented to be shed at a high frequency in contagious aerosols. On days 3 to 7 after infection of the donors, the 2 groups were housed in their respective chambers for 6 h and then in separate facilities, where samples were collected for testing by polymerase chain reaction and enzyme-linked immunosorbent assay over 14 d. Aerosol transmission was observed in 6 of the 20 replicates in the nonfiltered facility, whereas all pigs remained PRRSV-negative in the filtered facility; the difference was significant at P < 0.01. Thus, under the conditions of this study, the air-filtration system evaluated appeared to be highly effective at reducing aerosol transmission of PRRSV.     

Hemostatic changes in dogs with naturally occurring sepsis

Sepsis is a frequent source of morbidity and mortality in critically ill patients. The goal of this case control study was to measure hemostatic changes in dogs with naturally occurring sepsis. Blood was collected within 24 hours of admission from 20 dogs that fulfilled the criteria for sepsis. Sepsis was defined as histologic or microbiological confirmation of infection and 2 or more of the following criteria: hypo- or hyperthermia, tachycardia, tachypnea, or leukopenia, leukocytosis, or > 3% bands. Culture and sensitivities were performed on appropriate samples from all septic dogs. Twenty-eight control dogs were enrolled on the basis of normal results of physical examination, CBC, serum biochemistry, and coagulation profile. Plasma samples were analyzed for prothrombin time (PT), partial thromboplastin time (PTT), fibrin(ogen) degradation products (FDP), D-dimer (DD) concentrations, antithrombin (AT) activity, and protein C (PC) activity. Data were compared between groups by chi-square or independent t-tests. PC (P < .001) and AT (P < .001) activities were significantly lower in dogs with sepsis compared to controls. Dogs with sepsis had significantly higher PT (P = .007), PTT (P = .005), D-dimer (P = .005), and FDP (P = .001) compared to controls. Platelet counts were not significantly different between groups. Ten of the 20 septic dogs (50%) died, but no association was identified between any of the measured variables and outcome. These findings are consistent with previous studies in animals with experimentally induced disease and in clinical studies of humans. On the basis of these results, further investigation of the role of AT and PC in canine sepsis is warranted.     

Temporal trends in reproductive performance in Irish dairy herds and associated risk factors

: Irish dairy herd fertility has been declining since the 1980s. The extent, nature and causes of this decline in fertility and the current status of Irish dairy herd fertility were described. An increase in calving interval of approximately one day per year has been recorded. The principal components of this trend have been an increased incidence of postpartum endocrinopathies, reduced expression of oestrus and a fall in conception rate. Both submission rate and calving-to-service interval have increased slightly over time. Significant risk factors associated with these trends have been strain substitution within the Holstein-Friesian breed and single trait selection for milk production. Critically, these changes have been reflected in loss of body condition. Contributory factors included increased herd size and possibly increased use of DIYAI. The most recent Irish study showed that 48% of cows conceived to first service and 14% of cows were not pregnant at the end of the industry-average 15-week spring breeding season. However, the top quartile of herds achieved a first-service conception rate of 59%, illustrating the wide variation between herds. These phenotypic trends were attributed to both genetic and environmental factors and their interactions. Recent Irish dairy herd fertility performance falls short of the targets set for seasonal compact calving.     

Diagnostic considerations for evaluating nutritional problems in cattle.

The advances in testing methodology and in our understanding of the normal nutritional physiology of cattle have given veterinarians valuable tools to assess the nutritional status of cattle. By taking a representative number of appropriate samples, it is now relatively easy and inexpensive to determine the nutritional status of an individual or a group of animals. Also, the effectiveness of supplementation programs can be measured over time as part of an overall preventive medicine program.