Sero-epidemiological survey and risk factors associated with bovine brucellosis among slaughtered cattle in Nigeria

Bovine brucellosis is endemic in Nigeria; however, limited data exist on nationwide studies and risk factors associated with the disease. Using a cross-sectional sero-epidemiological survey, we determined the prevalence of and risk factors for brucellosis in slaughtered cattle in three geographical regions of Nigeria. Serum samples from randomly selected unvaccinated cattle slaughtered over a period of 3 years (between December 2010 and September 2013) from northern, southern and south-western Nigeria were tested for antibodies to Brucella abortus using the Rose Bengal test. Data associated with risk factors of brucellosis were analysed by Stata Version 12. In all, 8105 cattle were screened. An overall seroprevalence of 3.9% (315/8105) was recorded by the Rose Bengal test, with 3.8%, 3.4% and 4.0% from the northern, southern and south-western regions, respectively. Bivariate analysis showed that cattle screened in northern Nigeria were less likely to be seropositive for antibodies to Brucella spp. than those from south-western Nigeria (odds ratio = 0.94; 95% confidence interval: 0.73–1.22). However, logistic regression analysis revealed that breed ( p = 0.04) and sex ( p £ 0.0001) of cattle were statistically significant for seropositivity to Brucella spp. The study found that brucellosis was endemic at a low prevalence among slaughtered cattle in Nigeria, with sex and breed of cattle being significant risk factors. Considering the public health implications of brucellosis, we advocate coordinated surveillance for the disease among diverse cattle populations in Nigeria, as is carried out in most developed countries. Keywords: Bovine brucellosis, RBT, Epidemiology, Public Health, Nigeri

31st Annual Meeting and Associated Programs of the Society for Immunotherapy of Cancer (SITC 2016) : part two

Background The immunological escape of tumors represents one of the main ob- stacles to the treatment of malignancies. The blockade of PD-1 or CTLA-4 receptors represented a milestone in the history of immunotherapy. However, immune checkpoint inhibitors seem to be effective in specific cohorts of patients. It has been proposed that their efficacy relies on the presence of an immunological response. Thus, we hypothesized that disruption of the PD-L1/PD-1 axis would synergize with our oncolytic vaccine platform PeptiCRAd. Methods We used murine B16OVA in vivo tumor models and flow cytometry analysis to investigate the immunological background. Results First, we found that high-burden B16OVA tumors were refractory to combination immunotherapy. However, with a more aggressive schedule, tumors with a lower burden were more susceptible to the combination of PeptiCRAd and PD-L1 blockade. The therapy signifi- cantly increased the median survival of mice (Fig. 7). Interestingly, the reduced growth of contralaterally injected B16F10 cells sug- gested the presence of a long lasting immunological memory also against non-targeted antigens. Concerning the functional state of tumor infiltrating lymphocytes (TILs), we found that all the immune therapies would enhance the percentage of activated (PD-1pos TIM- 3neg) T lymphocytes and reduce the amount of exhausted (PD-1pos TIM-3pos) cells compared to placebo. As expected, we found that PeptiCRAd monotherapy could increase the number of antigen spe- cific CD8+ T cells compared to other treatments. However, only the combination with PD-L1 blockade could significantly increase the ra- tio between activated and exhausted pentamer positive cells (p= 0.0058), suggesting that by disrupting the PD-1/PD-L1 axis we could decrease the amount of dysfunctional antigen specific T cells. We ob- served that the anatomical location deeply influenced the state of CD4+ and CD8+ T lymphocytes. In fact, TIM-3 expression was in- creased by 2 fold on TILs compared to splenic and lymphoid T cells. In the CD8+ compartment, the expression of PD-1 on the surface seemed to be restricted to the tumor micro-environment, while CD4 + T cells had a high expression of PD-1 also in lymphoid organs. Interestingly, we found that the levels of PD-1 were significantly higher on CD8+ T cells than on CD4+ T cells into the tumor micro- environment (p < 0.0001). Conclusions In conclusion, we demonstrated that the efficacy of immune check- point inhibitors might be strongly enhanced by their combination with cancer vaccines. PeptiCRAd was able to increase the number of antigen-specific T cells and PD-L1 blockade prevented their exhaus- tion, resulting in long-lasting immunological memory and increased median survival

Co-infection of brucellosis and tuberculosis in slaughtered cattle in Ibadan, Nigeria: a case report

The authors present a case report on co-infection of brucellosis and tuberculosis in cattle slaughtered at the Bodija abattoir in Ibadan, Nigeria. Out of 32 animals that were seropositive for brucellosis using the Rose Bengal test, indirect enzyme-linked immunosorbent assay (ELISA) and competitive ELISA, six were also demonstrated as being infected with tuberculosis through mycobacterial culture. This is the first report of co-infection of brucellosis and tuberculosis in cattle slaughtered in Nigeria. There is a need for further studies to investigate this occurrence

Time-Resolved Fluorescent Resonance Energy Transfer Assay for Simple and Rapid Detection of Anti-Brucella Antibodies in Ruminant Serum Samples▿

Brucellosis is a globally significant zoonosis, the control of which is difficult and resource intensive. Serological tests form a vital part of a multifactorial approach to control and are often performed in large numbers. The aim of the present study was to develop a new assay to improve the efficiency, ease, and effectiveness of serological testing. An existing competitive enzyme-linked immunosorbent assay (cELISA) was adapted to a completely homogeneous time-resolved fluorescent resonance energy transfer (TR-FRET) assay. This was achieved by labeling an anti-Brucella monoclonal antibody with a long-lifetime donor fluorophore and Brucella smooth lipopolysaccharide with a compatible acceptor and optimizing the reading conditions. The assay was performed in a 96-well plate with a single 30-min incubation period and no separation (wash) steps and was concluded by a single plate-reading step. The performance of the assay was evaluated with a panel of serum samples from infected (n = 73) and uninfected (n = 480) sources and compared to the performance of the parent cELISA, an indirect ELISA (iELISA), and fluorescence polarization assay (FPA). The performance of the TR-FRET assay matched the performance of the iELISA, which had 100% diagnostic sensitivity and specificity, and surpassed the performance of the cELISA and the FPA. The results also demonstrated that the TR-FRET technique is effective with poor-quality serum samples from the field. To the knowledge of the authors, this is the first homogeneous TR-FRET assay to detect antibodies raised against an infectious disease. The technique appears to be sufficiently adaptable to meet the needs of many other similar testing requirements to identify infectious diseases

Knowledge and practices related to bovine brucellosis transmission amongst livestock workers in Yewa, south-western Nigeria

Brucellosis is an endemic disease in the animal population in Nigeria and of major public health importance, particularly amongst livestock workers who are ignorant of the risk of Brucella infection. Therefore, to gain insight into the knowledge and practices related to brucellosis transmission amongst livestock holders (LH) and livestock marketers (LM) in Yewa, an international livestock trading centre in south-western Nigeria, we conducted an interviewbased study using a cluster sampling technique. In all, a total of 157 respondents comprising 54 LH and 103 LM were interviewed. Two-thirds (69.5%) of the two groups had poor knowledge of brucellosis with no significant difference between them (p = 0.262). Furthermore, consumption of unpasteurised milk, uncooked meat and its products, co-habitation with animals, and poor hygiene were significant risk practices identified as possible means of transfer of Brucella infection from animals to humans amongst these livestock workers (p &lt; 0.05). In conclusion, our findings revealed that poor knowledge and practices related to the consumption of unpasteurised or unboiled dairy products, contaminated beef, and unhygienic practices are factors that will facilitate Brucella infections amongst livestock workers in Nigeria. Therefore, there is a need for more public health enlightenment programmes, as well as implementation of brucellosis control measures in the cattle populations

Serological investigation of bovine brucellosis in three cattle production systems in Yewa Division, south-western Nigeria

Limited data are available on the risk factors responsible for the occurrence of brucellosis amongst different cattle production systems in Nigeria despite its significant impact on livestock production. Consequently, a cross-sectional study was conducted to determine the prevalence of bovine brucellosis in three cattle production systems in Yewa Division of Ogun State, south-western Nigeria. A total of 279 blood samples (sedentary = 88; transhumance = 64; trade = 127) were examined for antibodies to Brucella sp. using the Rose Bengal test (RBT) and competitive enzyme-linked immunosorbent assay (cELISA). Overall, 24 (8.6%) and 16 (5.7%) of the animals tested seropositive for Brucella using RBT and cELISA, respectively. The herd seroprevalences based on RBT and cELISA were 31.6% and 15.8%, respectively. The results using cELISA reveal higher seroprevalence in the trade cattle (7.9%; confidence intervals [CI] = 3.2% – 12.6%) and those in a sedentary system (5.7%; CI = 0.9% – 10.5%) than in cattle kept under a transhumant management system (1.6%; CI = 1.5% – 4.7%). Age (> 3 years; p = 0.043) and breed (Djali; p = 0.038) were statistically significant for seropositivity to brucellosis based on cELISA, but sex (female, p = 0.234), production system (trade and sedentary; p = 0.208) or herd size (> 120; p = 0.359) was not. Since breeding stock is mostly sourced from trade and sedentary cattle, it is important that routine serological screening should be conducted before introducing any animal into an existing herd

Raloxifene and Desmethylarzoxifene Block Estrogen-Induced Malignant Transformation of Human Breast Epithelial Cells

There is association between exposure to estrogens and the development and progression of hormone-dependent gynecological cancers. Chemical carcinogenesis by catechol estrogens derived from oxidative metabolism is thought to contribute to breast cancer, yet exact mechanisms remain elusive. Malignant transformation was studied in MCF-10A human mammary epithelial cells, since estrogens are not proliferative in this cell line. The human and equine estrogen components of estrogen replacement therapy (ERT) and their catechol metabolites were studied, along with the influence of co-administration of selective estrogen receptor modulators (SERMs), raloxifene and desmethyl-arzoxifene (DMA), and histone deacetylase inhibitors. Transformation was induced by human estrogens, and selectively by the 4-OH catechol metabolite, and to a lesser extent by an equine estrogen metabolite. The observed estrogen-induced upregulation of CYP450 1B1 in estrogen receptor negative MCF-10A cells, was compatible with a causal role for 4-OH catechol estrogens, as was attenuated transformation by CYP450 inhibitors. Estrogen-induced malignant transformation was blocked by SERMs correlating with a reduction in formation of nucleobase catechol estrogen (NCE) adducts and formation of 8-oxo-dG. NCE adducts can be formed consequent to DNA abasic site formation, but NCE adducts were also observed on incubation of estrogen quinones with free nucleotides. These results suggest that NCE adducts may be a biomarker for cellular electrophilic stress, which together with 8-oxo-dG as a biomarker of oxidative stress correlate with malignant transformation induced by estrogen oxidative metabolites. The observed attenuation of transformation by SERMs correlated with these biomarkers and may also be of clinical significance in breast cancer chemoprevention