Effectiveness of vaccines and vaccination programs for the control of foot-and-mouth disease in Uganda, 2001–2010

Foot-and-mouth disease (FMD) is a highly contagious disease of cloven-hoofed animals. In Uganda, FMD outbreaks are mainly controlled by ring vaccination and restriction of animal movements. Vaccination stimulates immunity and prevents animals from developing clinical signs which include lameness, inappetence, and decreased production. Ring vaccination and restriction of animal movements have, however, not successfully controlled FMD in Uganda and outbreaks reoccur annually. The objective of this study was to review the use of FMD virus (FMDV) vaccines and assess the effectiveness of vaccination programs for controlling FMD in Uganda (2001–2010), using retrospective data. FMD vaccine distribution patterns in Uganda (2001–2010) matched occurrence of outbreaks with districts reporting the highest number of outbreaks also receiving the largest quantity of vaccines. This was possibly due to “fire brigade” response of vaccinating animals after outbreaks have been reported. On average, only 10.3 % of cattle within districts that reported outbreaks during the study period were vaccinated. The average minimum time between onset of outbreaks and vaccination was 7.5 weeks, while the annual cost of FMDV vaccines used ranged from US $58,000 to 1,088,820. Between 2001 and 2010, serotyping of FMD virus was done in only 9/121 FMD outbreaks, and there is no evidence that vaccine matching or vaccine potency tests have been done in Uganda. The probability of FMDV vaccine and outbreak mismatch, the delayed response to outbreaks through vaccination, and the high costs associated with importation of FMDV vaccines could be reduced if virus serotyping and subtyping as well as vaccine matching were regularly done, and the results were considered for vaccine manufacture.     

Immune dynamics following infection of avian macrophages and epithelial cells with typhoidal and non-typhoidal Salmonella enterica serovars; bacterial invasion and persistence, nitric oxide and oxygen production, differential host gene expression, NF-κB signalling and cell cytotoxicity

Poultry-derived food is a common source of infection of human with the non-host-adapted salmonellae while fowl typhoid and pullorum disease are serious diseases in poultry. Development of novel immune-based control strategies against Salmonella infection necessitates a better understanding of the host-pathogen interactions at the cellular level. Intestinal epithelial cells are the first line of defence against enteric infections and the role of macrophages is crucial in Salmonella infection and pathogenesis. While gene expression following Salmonella infection has been investigated, a comparison between different serovars has not been, as yet, extensively studied in poultry. In this study, chicken macrophage-like cells (HD11) and chick kidney epithelial cells (CKC) were used to study and compare the immune responses and mechanisms that develop after infection with different Salmonella serotypes. Salmonella serovars Typhimurium, Enteritidis, Hadar and Infantis showed a greater level of invasion and/or uptake characters when compared with S. Pullorum or S. Gallinarum. Nitrate and reactive oxygen species were greater in Salmonella-infected HD11 cells with the expression of iNOS and nuclear factor-κB by chicken macrophages infected with both systemic and broad host range serovars. HD11 cells revealed higher mRNA gene expression for CXCLi2, IL-6 and iNOS genes in response to S. Enteritidis infection when compared to S. Pullorum-infected cells. S. Typhimurium- and S. Hadar-infected HD11 showed higher gene expression for CXCLi2 versus S. Pullorum-infected cells. Higher mRNA gene expression levels of pro-inflammatory cytokine IL-6, chemokines CXCLi1 and CXCLi2 and iNOS genes were detected in S. Typhimurium- and S. Enteritidis-infected CKC followed by S. Hadar and S. Infantis while no significant changes were observed in S. Pullorum or S. Gallinarum-infected CKC.     

Bacterial toll-like receptor agonists induce sequential NF-κB-mediated leukotriene B4 and prostaglandin E2 production in chicken heterophils

Studies of the response of the primary avian polymorphonuclear leukocyte, the heterophil, to microbe associated molecular patterns (MAMPs) through toll-like receptors (TLR) has concentrated on the activation of the respiratory burst, release of intracellular granules, and the induction of cytokine and chemokine expression. Virtually no studies have been described on the role of lipid mediators, leukotrienes and prostaglandins, as effectors of the avian inflammatory response. We have previously shown that flagellin (FLG), the bacterial lipoprotein mimic palmitoly-3-cysteine-serine-lysine-4 (PAM), and unmethylated CpG motifs of bacteria DNA (CpG) are all potent activators of the avian innate immune system. In the present studies, we hypothesized that FLG, PAM, and CpG are also capable of eliciting the production of these lipid mediators of inflammation by avian heterophils. Compared to non-stimulated control heterophils, all three TLR agonists were potent inducers (3-5-fold increase) of a rapid production (30 min) of leukotriene B(4) (LTB(4)) followed by a later release (60-120 min) of prostaglandin (PGE(2)) by the heterophils. LTB(4) and PGE(2) production were derived from lipoxygenase-5 (5-LO) and cyclooxygenase-2 (COX-2) enzymatic activities, respectively, as the selective 5-LO (caffeic acid) and COX-2 (NS-398) inhibitors eliminated LTB(4) and PGE(2) production from the MAMP-stimulated heterophils. These results demonstrate that both the lipoxygenase and cycloxygenase pathways are operational in avian heterophils in response to bacterial MAMPs. Treatment of heterophils with either FLG, PAM, or CpG also induced a significant increase in DNA binding by NF-κB family members' p50, c-Rel, and RelB. Additionally, the production of LTB(4) and PGE(2) were inhibited following treatment of heterophils with the specific pharmacologic inhibitor of NF-κB (Bay 11-7086), thus suggesting that TLR pathway activation of NF-κB controls LTB(4) and PGE(2) production. This the first report of the production of lipid mediators of inflammation by avian heterophils in response to PAMPs. Since FLG, lipoproteins, and bacterial CpG DNA are abundant during bacterial infections, these data support their role in the inflammatory response mediated by avian heterophils.     

Involvement of the choroid plexus in the inflammatory response after acute spinal cord injury in dogs: An immunohistochemical study

The choroid plexus (CP) is increasingly recognized as an important contributor to central nervous system (CNS) inflammation by recruitment of inflammatory cells and release of inflammatory cytokines. Here we investigate the role of the CP epithelium (CPE) as a source of three pro-inflammatory molecules of potential importance in inflammation after acute spinal cord injury (SCI): IL-1β, TNF-α, and hsp70. Immunohistochemical (IHC) staining for these three proteins was performed on 4th ventricular CPE from 4 dogs euthanized 12-48h after spontaneous acute SCI, and from 4 neurologically normal dogs euthanized for other reasons. IHC staining was quantified using Aperio ImageScope software. IHC staining in the CPE of dogs with acute SCI was 2.2, 1.6 and 1.5 times higher than that of normal dogs, for IL-1β, TNF-α, and hsp70, respectively. Increases were statistically significant (p<0.1) for IL-1β and TNF-α, and closely approached significance for hsp70. These findings indicate that the CPE could serve as an important source of these inflammatory mediators after SCI. There was also an inverse correlation between IL-1β and hsp70 staining and duration of clinical signs in acute SCI, suggesting that increased expression of these proteins by the CPE may be of particular importance in the immediate-early inflammatory response after acute SCI.     

In vivo characterization of inflammatory biomarkers in swine and the impact of flunixin meglumine administration

Non-steroidal anti-inflammatory drugs (NSAID) are a family of chemicals that function to reduce pain, fever, and inflammation, and they are commonly used in people and animals for this purpose. Currently there are no NSAIDs approved for the management of inflammation in swine due to a lack of validated animal models and suitable biomarkers to assess efficacy. A previous in vitro study examining biomarkers of inflammation identified fourteen genes that were significantly altered in response to Escherichia coli lipopolysaccharide (LPS)-induced inflammation. In the present study, five of those fourteen genes were tested in vivo to determine if the same effects observed in vitro were also observed in vivo. Plasma levels of prostaglandin E(2) (PGE(2)), an essential mediator of fever and inflammation, were also determined. Two groups of swine were stimulated with LPS with the second group also treated with flunixin meglumine. Blood was collected at 0, 1, 3, 6, 8, 24, and 48h post LPS-stimulation. The RNA was extracted from the blood and quantitative real-time-PCR (qRT-PCR) was utilized to determine the expression patterns of CD1, CD4, serum amyloid A2 (SAA2), Caspase 1, and monocyte chemoattractant protein 1 (MCP-1). The LPS-stimulated animals demonstrated a statistically significant alteration in expression of SAA2 and CD1 at 3h post-stimulation. Flunixin meglumine treated animals' demonstrated reduced expression of CD1 in comparison to the LPS-stimulated swine at 24 and 48h post LPS-stimulation. Flunixin meglumine treated animals exhibited reduced expression of SAA2 at 48h post-stimulation compared to LPS-stimulated swine. Swine treated with LPS demonstrated statistically significant increases in plasma PGE(2) at 1h post-stimulation. Swine treated with flunixin meglumine had no increase in plasma PGE(2) levels at any time. These results demonstrate that PGE(2) production, along with two out of five genes (SAA2 and CD1) have the potential to serve as early biomarkers of inflammation as well as indicators of NSAID efficacy.     

Adenosine-5'-triphosphate release by Mannheimia haemolytica, lipopolysaccharide, and interleukin-1 stimulated bovine pulmonary epithelial cells

Mannheimia haemolytica, one of the agents associated with bovine respiratory disease complex, can cause severe lung pathology including the leakage of vascular products into the airways and alveoli. Previous work by this laboratory has demonstrated that bovine lung endothelial and epithelial cells undergo dramatic permeability increases when exposed to adenosine-5'-triphosphate (ATP). Therefore, we wanted to determine if ATP levels were elevated in bronchoalveolar lavage (BAL) samples from calves experimentally infected with M. haemolytica. In addition, cultured bovine pulmonary epithelial (BPE) cells were stimulated with heat-killed and live M. haemolytica bacteria, lipopolysaccharide (LPS), lipoteichoic acid (LTA), interleukin-1 (IL-1), and zymosan activated plasma (ZAP) to determine whether they might release extracellular ATP during in vitro infection. Calves experimentally exposed to M. haemolytica had an approximately 2-fold higher level of ATP in their BAL samples compared to control. BPE cells exposed to increasing numbers of heat-killed or live M. haemolytica had significantly increased levels of ATP release as compared to time-matched controls. Finally, BPE cells treated with several concentrations of LPS and IL-1 had increases in ATP release as compared to time-matched controls. This increase appeared to be a result of active ATP secretion by the cells, as cell viability was similar between treated and non-treated cells. Neither ZAP nor LTA induced any ATP release by the cells. In conclusion, ATP levels are elevated in lung secretions from calves infected with M. haemolytica. In addition, lung epithelial cells can actively release ATP when exposed to heat-killed or live M. haemolytica, LPS or IL-1.     

Experimental infection by capillary tube feeding of Rhipicephalus sanguineus with Bartonella vinsonii subspecies berkhoffii

It has been speculated that ticks may serve as vectors of Bartonella species. Circumstantial, clinical, epidemiological and serological evidence suggest that B. vinsonii subspecies berkhoffii (B. v. berkhoffii) might be transmitted by Rhipicephalus sanguineus. The purpose of the present study was to determine whether adult R. sanguineus ticks can be infected with a B. v. berkhoffii genotype II isolate via capillary tube feeding and whether the infection can then be transmitted from adult females to their eggs via trans-ovarial transmission. Furthermore, tick fecal material was also collected and screened as a possible source of infectious inoculum for canine infections. B. v. berkhoffii DNA was detected in 50% (7 of 14) of females that did not oviposit and in 14.3% (2 of 14) of female ticks that laid eggs, but not detected in egg clutches (100 eggs/female). DNA was also detected in tick feces collected on days 2 through 6 post-capillary tube feeding, however, dogs (n=3) did not become bacteremic or seroconvert when inoculated with tick fecal material. Therefore, trans-ovarial transmission of B. v. berkhoffii by R. sanguineus is unlikely, but further studies are needed to determine if tick fecal material can serve as a source of infection to canines.