An investigation into the role of Chlamydophila spp. in bovine upper respiratory tract disease

An outbreak of upper respiratory tract disease was investigated in a group of 17 housed home-bred calves on a mixed dairy, beef and sheep farm in Devon. Conjunctival swabs were collected and tested for Chlamydophila spp. DNA using a PCR test that detects Chlamydophila abortus and Chlamydophila psittaci. Six of the calves tested gave a positive result. Further epidemiological observations and laboratory testing indicated that the adult dairy cows, from which the affected calves originated, were the most likely source of infection.     

Trends in the incidence of tuberculosis in possums and livestock, associated with differing control intensities applied to possum populations

AIM: To determine the trap-catch index (an estimate of abundance) of brushtail possum (Trichosurus vulpecula) populations infected with bovine tuberculosis (Tb; Mycobacterium bovis) that must be achieved, and the length of time such an index must be maintained, for Tb to be eliminated from possum populations and adjacent livestock. METHODS: Between 1997-1998 and 2000-2001, trap-catch surveys of possum populations naturally infected with Tb and subjected to population-control measures were undertaken at four forest sites and two farmland sites. At the same time, possum carcasses were collected at these sites and their Tb status determined, and all contiguous cattle and deer herds were Tb tested and abattoir slaughter data for these herds were interrogated. RESULTS: Trap-catch surveys indicated that numbers of possums on the farmland sites surveyed were usually very low and well below the control targets set (i.e. a 5% trap catch or approximately 0.5-1 possum/ha) for the study. In contrast, trap-catch surveys undertaken in forest sites indicated possum numbers were more variable, and often recovered rapidly from control operations to exceed control targets within 1-3 years. The annual rate of recovery of possum populations in half of the forest population surveys undertaken exceeded published intrinsic rates of increase for possums. The overall prevalence of Tb in possum populations was < or =1.9% at 5/6 sites, and was 6.5% at the sixth site. Juvenile possums infected with Tb were trapped within but near the edge of control zones and appeared to represent an immigrant source of infection. Mature infected possums survived control operations apparently by having home ranges in uncontrolled patches within control areas. Infection in possums appeared to be eliminated from one study site by the intensive control undertaken, but elimination at other sites appeared less likely. Levels of Tb in livestock on or adjacent to the study sites fell by at least 50% during the study, and cattle in one area tested clear for the first time in 20 years. CONCLUSIONS: Initial control of possums in forest appeared to achieve national control targets set by the Animal Health Board (AHB), despite trap-catch data often providing misleading population estimates. Such targets were often exceeded within 1-3 years. By comparison, possum control on farmland appeared to maintain populations at very low levels, while control on forest margins maintained populations at intermediate levels. Control was least effective in deep forest where human access was most difficult. Intensive population control measures appeared to have led to a reduced incidence of Tb in livestock at 3/4 sites, and elimination of Tb in livestock at one site. This result supports modelling studies that predict the eradication of Tb from possums through ongoing intensive control and may explain the lower success achieved with earlier less-intensive possum control.     

Echinococcus multilocularis in Austrian foxes from 1991 until 2004

The prevalence rates of Echinococcus multilocularis in foxes (n=5600) evaluated in several Austrian surveys conducted between 1991 and 2004 were analysed for spatial and temporal differences. Data from early studies (1993-1997) in which the intestinal scraping technique (IST) was utilized were compared with data from recent (1999-2004) investigations, which made use of the shaking in a vessel technique (SVT), and it was assessed whether or not the infection rates of Austrian foxes had increased between the investigated intervals. In total, data from 85 districts are presented and both the retrospective and recent data are available from 39 of these districts. A Bayesian hierarchical model of parasite prevalences is presented which (i) accounts for differences in the sensitivity of IST and SVT, (ii) incorporates spatial auto-correlation between neighbouring districts, (iii) investigates the possibility of a temporal shift in the infection status of foxes, and (iv) quantifies uncertainty at each level of the model. The national average prevalence rates in the mid-1990s and at the turn of the millennium were 2.4% (95% confidence intervals 1.1-4.8) and 3.9% (95% confidence intervals 1.5-8.4) respectively. Above average prevalence rates were observed in the western and the northern parts of the country. Evidence is also presented for a temporal augmentation of the prevalence rates in some districts in the northern and eastern parts of the country. These findings are in concordance with several investigations in other European states where both newly emerged areas and elevated levels of transmission in existing endemic areas have been found. None of the districts investigated here showed significant evidence of a drop in prevalence.     

Detection of chlamydiae in boar semen and genital tracts

Chlamydiae cause abortion and reproductive disorders in sows. Although organisms can infect the male genital tract, little is known about the disease situation in boars. Hence, we examined the prevalence of chlamydial infection in semen and genital tracts of boars. Samples collected from Swiss boars (group A: n=42), and boars from Germany (group B: n=39) were examined by bacteriology, LPS-ELISA, immunohistochemistry (IHC) and polymerase chain reaction (PCR). The latter methodology involved use of three PCR assays including 16Sig rDNA, IGS-S (intergenic spacer 16S/23S-Short) and IGS-L (intergenic spacer 16S/23S-Long) PCR for comparison methods. PCR sensitivity and the presence of potential PCR inhibitors were determined by spiking semen with Chlamydophila (Cp.) abortus DNA. Detection limits of the 16Sig and IGS-S PCR were 10 templates, while the IGS-L PCR was less sensitive (100 templates). Of 25 semen samples that were collected from group A, one semen sample was positive for Cp. psittaci and two were positive for Chlamydia-like organisms by 16Sig PCR. Screening of sera from Swiss boars revealed three animals with positive reactions in the LPS-ELISA, although we failed to detect chlamydiae within organs of these or sera-negative animals by IHC or IGS-S PCR. In group B, 10 ejaculates were positive for Chlamydia (C.) suis and two were positive for Chlamydia-like organisms by 16S PCR. The identification of DNA from Chlamydia-like organisms in semen from both groups of boars was surprising and a role for these bacteria in reproductive diseases requires further assessment. In conclusion, the prevalence of chlamydial infection was low in group A animals indicating that venereal transmission may not be significant for Chlamydia-associated reproductive diseases in pigs, although rare cases may occur.     

Developments and hurdles in generating vaccines for controlling helminth parasites of grazing ruminants

As a direct consequence of rising drug resistance among common nematodes of grazing animals, efforts toward state-of-the-art vaccine development have clearly intensified in recent years, fuelled primarily by the advent of newer technologies in gene discovery, by advancements in antigen identification, characterisation and production. In this regard, it is appropriate to review progress that has been made in generating helminth vaccines and in particular, vaccines against common nematodes of production animals for consumption. In like manner, it is prudent to evaluate barriers that have hindered progress in the past and continue to present obstacles that must be solved when utilizing and depending on host immunity to attenuate parasitic infections.     

Response of Trypanosoma vivax and Trypanosoma congolense in zebu cattle in North Cameroon to prophylactic treatment with two formulations of isometamidium

We tested the efficacy of two formulations of isometamidium in a tsetse-infested farm in North Cameroon from 20 August 2000 to 5 January 2001. A total of 90 adult cattle were used in three groups of 30 each corresponding to two treated and one untreated control. Drug efficacies were evaluated in terms of reduction of parasite incidence in the host's blood, maintenance of packed-cell volume (PCV) and weight gains. Both drugs reduced the incidence of parasites even though re-infections 2 weeks after treatment were common. PCV values were similar in both treated groups but higher than in the untreated control. Body weight changes followed a similar trend with the control losing weight from a mean of 427+/-119kg at the beginning to 398+/-93kg in 4 months. Weights increased from 375+/-76 and 396+/-110 to 396+/-69 and 418+/-112kg in the Veridium and Trypamidium groups, respectively. Efficacy was similar between the two formulations of isometamidium in the prophylaxis of bovine trypanosomosis. However, the presence of parasites in some animals barely 2 weeks after treatment suggested that either infections were not cleared or residual drug effects were not sufficient to prevent re-infections.     

Regulation by Jun N-terminal kinase/stress activated protein kinase of cytokine expression in Mycobacterium avium subsp paratuberculosis-infected bovine monocytes

OBJECTIVE: To evaluate activation of Jun N-terminal kinase/stress-activated protein kinase (JNK/SAPK) pathway in bovine monocytes after incubation with Mycobacterium avium subsp paratuberculosis (Mptb) organisms. SAMPLE POPULATION: Bovine monocytes obtained from 4 healthy adult Holstein dairy cows. PROCEDURES: Bovine monocytes were incubated with Mptb organisms with or without a specific inhibitor of the JNK/SAPK pathway (SP600125) for 2, 6, 24, or 72 hours. Expression of interleukin (IL)-1beta, IL-10, IL-12, IL-18; transforming growth factor-beta (TGF-beta); and tumor necrosis factor-alpha (TNF-alpha) and the capacity of Mptb-infected monocytes to acidify phagosomes and kill Mptb organisms were evaluated. Phosphorylation status of JNK/SAPK was evaluated at 10, 30, and 60 minutes after Mptb incubation. RESULTS: Compared with uninfected control monocytes, Mptb-infected monocytes had increased expression of IL-10 at 2 and 6 hours after incubation and had increased expression of TNF-alpha, IL-1beta, IL-18, and TGF-beta at 2, 4, and 6 hours. Additionally, Mptb-infected monocytes had increased expression of IL-12 at 6 and 24 hours. Addition of SP600125 (specific chemical inhibitor of JNK/SAPK) resulted in a decrease in TNF-alpha expression at 2, 6, and 24 hours, compared with untreated Mptb-infected cells. Addition of SP600125 resulted in a decrease in TGF-beta expression at 24 hours and an increase in IL-18 expression at 6 hours. Addition of SP600125 failed to alter phagosome acidification but did enhance the capacity of monocytes to kill Mptb organisms. CONCLUSIONS AND CLINICAL RELEVANCE: Activation of JNK/SAPK may be an important mechanism used by Mptb to regulate cytokine expression in bovine monocytes for survival and to alter inflammatory and immune responses.     

Sex steroids level in blood plasma and ovarian follicles of the chimeric chicken

The study was performed to determine the hormonal status of mature germline chimeras obtained by blastodermal cell transfer from chicken embryos of a donor breed [Green-legged Partridgelike breed (GP) x Araucana (AR)] to those of a recipient breed [White Leghorn (WL)] being at the same stage of embryonic development. The egg-laying chimeras and WL hens (control) of the same age were used in the experiment. At first, blood samples were taken from each bird at 0.5, 5, 12.5 and 18.5 h following oviposition. Subsequently, the chimeras and the WL hens were decapitated 1-2 h after ovulation. A stroma and the following follicles were isolated from the ovary: white normal (1-4, 4-6 and 6-8 mm), white atretic and yellow preovulatory follicles (F4-F1). Sex hormones, progesterone (P4), testosterone (T) and oestradiol (E2) in blood plasma and ovarian follicles were determined radioimmunologically. The activity of the 3beta-hydroxysteroid dehydrogenase (3beta-HSD) in the granulosa and theca layers of the follicles was analysed histochemically. In chimeric chickens, a higher level of T in blood plasma during the ovulatory cycle was noticed. However, in the stroma, white prehierarchical and medium-size preovulatory ovarian follicles the level of T was significantly lower. With respect to E2, its elevated levels were found both in blood and in the ovarian follicles. There were no significant differences in P4 concentrations in blood plasma while in ovarian follicles a higher level was observed only in white 6-8 mm follicles. 3beta-HSD activity in granulosa and theca layers of the ovarian follicles in chimeras was not different from that in the WL hens. In conclusion, the results obtained indicate that germline chimeras exhibit significant alterations in sex hormone levels in the ovary and blood plasma, which in turn may affect their reproductive abilities.     

Circulating ghrelin concentrations fluctuate relative to nutritional status and influence feeding behavior in cattle

The objective of these experiments was to establish the relationship of plasma ghrelin concentrations with feed intake and hormones indicative of nutritional state of cattle. In Exp.1, 4 steers (BW 450 +/- 14.3 kg) were used in a crossover design to compare plasma ghrelin concentrations of feed-deprived steers with those of steers allowed to consume feed and to establish the relationship of plasma ghrelin concentrations with those of GH, insulin (INS), glucose (GLU), and NEFA. After adaptation to a once-daily feed offering (0800), 2 steers continued the once-daily feeding schedule (FED), whereas feed was withheld from the other 2 steers (FAST). Serial blood samples were collected via indwelling jugular catheter from times equivalent to 22 h through 48 h of feed deprivation. Average plasma ghrelin concentrations were greater (P < 0.001) in FAST compared with FED (690 and 123 +/- 6.5 pg/mL) steers. Average plasma ghrelin concentrations for FED steers prefeeding were elevated (P < 0.001) when compared with those postfeeding (174 and 102 +/- 4.2 pg/mL, respectively). Average plasma GH concentration was elevated (P < 0.05) for FAST steers compared with FED steers. Plasma GLU concentrations were not different; however, for FAST steers, NEFA concentrations were elevated (P < 0.001) and INS concentrations were decreased (P < 0.001). In Exp. 2, 4 steers (BW 416 +/- 17.2 kg) were used in a crossover design to determine the effects of i.v. injection of bovine ghrelin (bGR) on plasma GH, INS, GLU, and NEFA concentrations; length of time spent eating; and DMI. Steers were offered feed once daily (0800). Serial blood samples were collected from steers via indwelling jugular catheter. Saline or bGR was injected via jugular catheter at 1200 and 1400. A dosage of 0.08 microg/kg of BW bGR was used to achieve a plasma ghrelin concentration similar to the physiological concentration measured in a FAST steer in Exp. 1 (1,000 pg/mL). Injection of bGR resulted in elevated (P < 0.005) plasma GH concentrations after the 1200 but not the 1400 injection. Plasma INS, GLU, and NEFA concentrations were not affected by bGR injection. For the combined 1-h periods postinjection, length of time spent eating was greater (P = 0.02) and DMI tended to be increased (P = 0.06) for bGR steers. These data are consistent with the hypothesis that ghrelin serves as a metabolic signal for feed intake or energy balance in ruminants.     

American foulbrood of the honey bee: Occurrence and distribution of different genotypes of Paenibacillus larvae in the administrative district of Arnsberg (North Rhine‐Westphalia)

Between March 2003 and October 2004, Paenibacillus larvae, the aetiological agent of American foulbrood disease of the honey bee, was isolated from broodcombs and honey samples of 54 apiaries in the administrative district of Arnsberg (North Rhine‐Westphalia, Germany). Genotyping of 176 P. larvae isolates with repetitive element polymerase chain reaction fingerprinting (rep‐PCR) using BOX A1R and MBO REP1 primers revealed five different genotypes (AB, Ab, ab, aß, AБ). In samples of three apiaries, more than one genotype was detected. A combination of two genotypes was isolated from honey samples of the same hive two times (ab/aß and Ab/ab). The five genotypes were not randomly distributed in the district, but revealed a certain geographical clustering. Possible factors with impact on the genotype diversity and the distribution pattern are discussed.