TB diagnosis in non-human primates: comparison of two interferon-gamma assays and the skin test for identification of Mycobacterium tuberculosis infection

To examine the effect of recombinant bovine interferon-gamma (rbIFN-gamma) on cattle persistently infected with bovine leukemia virus (BLV), BLV-infected cattle were inoculated intraperitoneally with IFN-gamma. All cattle were febrile after inoculation with IFN-gamma and then recovered within 48 h. Flow cytometric analysis showed that the numbers of CD4+ and CD8+ T cells were decreased for 2-3 days and then their numbers were recovered. The number of gammadelta T cells increased after the fever. In contrast, the number of IgM+ lymphocytes remained low for about 1 week. Moreover, the numbers of syncytia produced by peripheral blood lymphocytes decreased and remained low compared to that before IFN-gamma administration. These results suggest that IFN-gamma induces the up-regulation of gammadelta T cells, decreases the number of IgM+ lymphocytes and suppresses the growth of BLV in BLV-infected cattle in vivo.     

Feline calicivirus

Feline calicivirus (FCV) is an important and highly prevalent pathogen of cats. It belongs to the family Caliciviridae which includes other significant pathogens of man and animals. As an RNA virus, high polymerase error rates convey upon FCV a high genome plasticity, and allow the virus to respond rapidly to environmental selection pressures. This makes the virus very adaptable and has important implications for clinical disease and its control. Being genetically diverse, FCV is associated with a range of clinical syndromes from inapparent infections to relatively mild oral and upper respiratory tract disease with or without acute lameness. More recently, highly virulent forms of the virus have emerged associated with a systemic infection that is frequently fatal. A proportion of FCV infected cats that recover from acute disease, remain persistently infected. In such cats, virus evolution is believed to help the virus to evade the host immune response. Such long-term carriers may only represent a minority of the feline population but are likely to be crucial to the epidemiology of the virus. Vaccination against FCV has been available for many years and has effectively reduced the incidence of clinical disease. However, the vaccines do not prevent infection and vaccinated cats can still become persistently infected. In addition, FCV strain variability means that not all strains are protected against equally. Much progress has been made in understanding the biology and pathogenesis of this important feline virus. Challenges for the future will necessarily focus on how to control the variability of this virus particularly in relation to emerging virulent strains and vaccination.     

Molecular and serologic characterization, pathogenicity, and protection studies with infectious bronchitis virus field isolates from California

In this study, we characterized three variant infectious bronchitis virus (IBV) strains isolated in 2003 and 2004 from broiler chickens in California and compared them to previously isolated California variant viruses and to common vaccine serotypes used in the United States. We conducted genetic, serologic, and pathogenicity studies on all three isolates, then tested different vaccines against one of the viruses. Genetically the three variant IBV strains, designated CA557/03, CA706/03, and CA1737/04, were not related to each other. GenBank BLAST database search and phylogenetic analysis of the hypervariable region of the S1 subunit of the spike gene to determine the most closely related viruses to the three variants showed the CA557/03 variant to be 81.8% similar to the CAV/CA56b/91 whereas the CA706/03 and CA1737/04 variant viruses were only distantly related to Dutch/D1466/81 (72.2%), a vaccine strain used in Europe, and Korea/K142/02 (72.7%), a Korean field isolate, respectively. Cross virus-neutralization testing showed that none of the 2003-04 California IBV variant viruses were serologically related to each other or to Ark, Conn, or Mass vaccine strains. In addition the CA1737/04 isolate was also tested against DE072 and found not to be serologically related. All three variant viruses were pathogenic in 1-wk-old broilers and vaccination with Mass/Conn followed by Holland/Conn provided 80% protection against the CA1737/04 virus. The 2003-04 California variant viruses were not compared with variants isolated in California during 1970s and 1980s because, to our knowledge, no genetic information is available and those viruses are no longer obtainable. This study shows that the CA557/03 virus was distantly related to the CAV-type viruses isolated in California in the early 1990s, but that none of the 2003-04 viruses were similar genetically or serologically to the CAL99-type viruses, indicating that new IBV variants continue to emerge and cause disease in commercial chickens in California.     

Comparing the detection of exposure to Ehrlichia ruminantium infection on a heartwater-endemic farm by the pCS20 polymerase chain reaction assay and an indirect MAP1-B enzyme linked immunosorbent assay

Detection of heartwater is not always easy especially because all the serological assays so far available either have poor sensitivity or specificity. The indirect MAP-1B ELISA has been reported to be the most specific test for heartwater, although it does also detect antibodies to some closely related ehrlichial agents. This study was undertaken to compare two methods for the detection of heartwater infection caused by the ehrlichial agent Ehrlichia (Cowdria) ruminantium. Fifteen cattle on a heartwater-endemic farm infested with high numbers of Amblyomma hebraeum ticks, and hence exposure to E. ruminantium infection were monitored over an 8-week period by pCS20 PCR and an indirect MAP-1B ELISA. Infection was detected by pCS20 PCR in most animals with the highest number of positives (60%) in week 6 of the study. Similarly, exposure to E. ruminantium was detected by indirect MAP-1B ELISA in some animals, with the highest number of seropositives (27%) at weeks 2-6 of the study. The data demonstrated a fluctuating rickettsaemia in cattle in a heartwater-endemic area. Comparison of the two tests indicated that the pCS20 PCR assay was more reliable because it detected more infections than the indirect MAP-1B ELISA and would therefore be the method of choice for detection of E. ruminantium infection.     

Socio-economic factors influencing control of vector-borne diseases in the pastoralist system of south western Uganda

Research in control of tick-borne diseases and trypanosomosis, and their vectors, namely, ticks and tsetse flies respectively, has been on going for decades. However, very little attention has been paid to the socio-economic factors that are likely to influence the outcome of the interventions in the control of these diseases. Thus, this study was designed to investigate these factors, mainly the intra-household factors influencing decision-making in the control of Vector-borne diseases in the pastoralist areas of Uganda. These factors included: indigenous technical knowledge, household economic factors, and gender. Both qualitative and quantitative methods were used in the collection and analysis of data. The tools used for data collection included among others, participatory learning and action (PLA), and Case studies. The findings included the following: In pastoralist households, a big proportion of the household budget was allocated to vector-borne diseases control. In the male-headed households, men dominated decision-making on vector-borne diseases control, although the goals and priorities of men and women in these households were not the same. Also, vector-borne disease control was predominantly by use of modern veterinary drugs, and pastoralists treated sick cattle by themselves even in situations where there were veterinary personnel.     

Validating an empiric sulfadiazine–trimethoprim dosage regimen for treatment of Escherichia coli and Staphylococcus delphini infections in mink (Neovison vison)

Antimicrobial agents are used extensively off‐label in mink, as almost no agents are registered for this animal species. Pharmacokinetic (PK) and pharmacodynamic (PD) data are required to determine antimicrobial dosages specifically targeting mink bacterial pathogens. The aims of this study were to assess, in a PKPD framework, the empirical dosage regimen for a combination of trimethoprim (TMP) and sulfadiazine (SDZ) in mink, and secondarily to produce data for future setting of clinical breakpoints. TMP and SDZ PK parameters were obtained experimentally in 22 minks following IV or oral administration of TMP/SDZ (30 mg/kg, i.e. 5 mg/kg TMP and 25 mg/kg SDZ). fAUC/MIC with a target value of 24 hr was selected as the PKPD index predictive of TMP/SDZ efficacy. Using a modeling approach, PKPD cutoffs for TMP and SDZ were determined as 0.062 and 16 mg/L, respectively. By incorporating an anticipated potentiation effect of SDZ on TMP against Escherichia coli and Staphylococcus delphini, the PKPD cutoff of TMP was revised to 0.312 mg/L, which is above the tentative epidemiological cutoffs (TECOFF) for these species. The current empirical TMP/SDZ dosage regimen (30 mg/kg, PO, once daily) therefore appears adequate for treatment of wild‐type E. coli and S. delphini infections in mink.     

Physiological Predictors in Calves of Dairy Breeds: Part 1. Genetic Parameters of Basal and Induced Growth Hormone Secretion

Abstract

Genetic variation in plasma growth hormone (GH) concentration before and after GRF (growth hormone releasing factor) stimulation was studied in young bulls (N=284) and heifers (N=212), the progeny of 53 sires of four dairy and dual-purpose breeds (Danish Jersey, Red Dane, Danish Friesian and Danish Red and White). Male and female calves were reared, fed and tested on separate experimental stations; thus sex, station and feeding were completely confounded effects. The animals were tested at about 9 months of age, after a 24 h fast. GH concentration was measured in serial plasma samples collected for 1 h before and 1 h following intravenous administration of 2.0 ug synthetic GRF(1–29)NH₂/kg live weight^>0.75. Prior to statistical analysis, concentrations were log_e-transformed. Response variables were BASELINE (mean GH in -15, -5 and 0 min samples) and PEAK (mean GH in 10, 15 and 20 min samples). A statistical model taking at least three generations of ancestral relationships into account was used to estimate variance and covariance components for traits in male and female calves by use of restricted maximum likelihood methods.

Heritability of BASELINE was low (0.04 ± 0.12) in males but high in females (0.60 ± 0.16). The heritability of PEAK was high in both sexes (males, 0.42 ± 0.16; females, 0.60 ± 0.16). Genetic correlations between the same trait measured in males and females were low for BASELINE (r _g = 0.32±0.55) but high for PEAK (r _g = 0.82±0.15). Within sex, BASELINE and PEAK were both highly genetically correlated (males, r _g=0.62 ± 0.40; females, r _g= 1.00 ± 0.07).

We conclude that growth hormone concentration is a highly heritable trait in juvenile cattle of both sexes, and that GRF stimulation is benificial to the uncovering of genetic differences among animals.     

The Effect of Inoculation Treatment and Long-term Application of Moisture on Fusarium Head Blight Symptoms and Deoxynivalenol Contamination in Wheat Grains

Fusarium head blight (FHB) is an important disease of wheat, which can result in the contamination of grains with mycotoxins such as deoxynivalenol (DON). Artificial inoculation of flowering ears with conidial suspensions is widely used to study FHB diseases. Our goal was to compare four inoculation treatments in which a conidial suspension was sprayed on flowering ears and to study the effect of the application of moisture during kernel setting and filling with a mist-irrigation system. Ten wheat genotypes were inoculated with a DON-producing Fusarium culmorum strain. Inoculation treatments varied in time of application of the inoculum (morning or evening) and in the method of controlling humidity during inoculation (bagging or mist irrigation). A wet season was simulated with a mist-irrigation system, keeping the crop canopy wet for at least 26 days after flowering. The severity of FHB symptoms (area under disease progress curve (AUDPC)), yield loss and DON contamination in the grains were determined. AUDPC data obtained with the different inoculation treatments were highly correlated (r=0.85–0.95). Mist irrigation after inoculation resulted in a higher mean disease severity, but in a overall lower toxin contamination as compared to the non-irrigated treatments. Genotypic differences in DON accumulation were present: for one wheat line toxin contamination significantly increased when irrigated, while two genotypes accumulated significantly less toxin. The closest relationships (r=0.73–0.89) between the visual symptoms and the DON content were obtained under moderate mean infection pressure. This relation between visual symptoms and the DON content deteriorated at higher infection levels.     

The presence of double-stranded RNAs in <Emphasis Type="Italic">Alternaria alternata</Emphasis> Japanese pear pathotype is associated with morphological changes

Strains of the Japanese pear pathotype of Alternaria alternata were screened for double-stranded RNAs (dsRNAs). Four strains had several dsRNAs; strain N18 was associated with several dsRNAs and had impaired growth phenotypes such as irregular mycelium and abnormal pigmentation. We isolated dsRNA-cured isolates from strain N18 by single-conidium isolation. The dsRNA-cured isolates had recovered normal growth and pigmentation. Enlarged vesicles were observed in mycelial cells of the original dsRNA-carrying N18 strain. DAPI nuclear staining revealed regression of the nuclei in dsRNA-carrying N18 cells. These results indicate that the dsRNAs might have negative effects, such as apoptosis-like cell death, on the host fungus.