Molecular diagnosis of the tick-borne pathogen Anaplasma marginale in cattle blood samples from Nigeria using qPCR.

Tick-borne diseases (TBDs) are some of the most important animal health and management problems in Africa, including Nigeria. This study aims to determine the prevalence of an important TBD, anaplasmosis, in a North-central region of Nigeria. Blood samples were collected from cattle and stored on Whatman FTA® cards. Information on village, age and sex associated with each cattle was also recorded. The packed red blood cell volume (PCV) for each blood sample was determined. After DNA extraction, pathogen presence was evaluated by TaqMan® based qPCR of which 75.9 % of the cattle tested positive for Anaplasma marginale. Statistical analysis revealed that the presence of A. marginale infection differed significantly between cattle age groups. However, there was no significant difference in the prevalence of this pathogen between the sexes or among cattle grouped by PCV level. Finally, using a highly sensitive molecular method our pioneer study contributes to the improvement of the current knowledge regarding tick-borne pathogens that seriously affect animal health in specific areas of Nigeria.

Reassortant Avian Influenza A(H9N2) viruses in chickens in retail poultry shops, Pakistan, 2009-2010.

Phylogenetic analysis of influenza viruses collected during December 2009-February 2010 from chickens in live poultry retail shops in Lahore, Pakistan, showed influenza A(H9N2) lineage polymerase and nonstructural genes generate through inter- and intrasubtypic reassortments. Many amino acid signatures observed were characteristic of human isolates; hence, their circulation could enhance inter- or intrasubtypic reassortment.

Associations between trematode infections in cattle and freshwater snails in highland and lowland areas of Iringa Rural District, Tanzania.

The epidemiology of trematode infections in cattle was investigated within highland and lowland areas of Iringa Rural District, in southern Tanzania. Fecal samples were collected from 450 cattle in 15 villages at altitudes ranging from 696 to 1800 m above the sea level. Freshwater snails were collected from selected water bodies and screened for emergence of cercariae. The infection rates in cattle were Fasciola gigantica 28·2%, paramphistomes 62·8% and Schistosoma bovis 4·8%. Notably, prevalence of trematode infections in cattle was much higher in highland (altitude > 1500 m) as compared with lowland (altitude < 1500 m) areas and was statistically significant (P-value = 0·000) for F. gigantica and paramphistomes but not for S. bovis. The snails collected included Lymnaea natalensis, Bulinus africanus, Bulinus tropicus, Bulinus forskali, Biomphalaria pfeifferi, Melanoides tuberculata and Bellamya constricta with a greater proportion of highland (75%) than lowland (36%) water bodies harbouring snails. Altitude is a major factor shaping the epidemiology of F. gigantica and paramphistomes infections in cattle in Iringa Rural District with greater emphasis upon control needed in highland areas.

Online tools for teaching evidence-based veterinary medicine.

Evidence-based veterinary medicine (EBVM) is of interest and relevance to veterinary practitioners. Consequently, veterinary schools take responsibility for teaching students how to appraise scientific articles and for equipping them with the skills needed to obtain and evaluate the best evidence and to apply this approach to their own cases. As part of our farm animal clinical rotation, we train students in qualitative and quantitative EBVM methods using an e-learning environment, online teaching materials, a wiki (a Web site that allows its users to edit its content via a Web browser), and face-to-face tutorials that support learning. Students working in small groups use a wiki to record details of the history, clinical presentation, diagnostic tests, herd data, and management plans for their chosen farm animal clinical cases. Using a standardized patient, intervention, comparison, and outcome (PICO) format, each group formulates a patient question based on either a proposed intervention or diagnostic procedure for the case and conducts an online scientific literature database search. The students appraise the articles retrieved using EBVM approaches and record the information in the wiki. The summation of this body of work, the group's critically appraised topic (CAT), includes the original PICO, a standardized table of the scientific evidence for the effectiveness of the intervention or diagnostic procedure, a summary statement in the form of a clinical bottom line, and their reflections upon the CAT. At the end of the rotation, students take part in a structured "CAT Club" where they present and discuss their findings with fellow students and clinicians.

Herd prevalence of bovine brucellosis and analysis of risk factors in cattle in urban and peri-urban areas of the Kampala economic zone, Uganda.

BACKGROUND: Human brucellosis has been found to be prevalent in the urban areas of Kampala, the capital city of Uganda. A cross-sectional study was designed to generate precise information on the prevalence of brucellosis in cattle and risk factors for the disease in its urban and peri-urban dairy farming systems. RESULTS: The adjusted herd prevalence of brucellosis was 6.5% (11/177, 95% CI: 3.6%-10.0%) and the adjusted individual animal prevalence was 5.0% (21/423, 95% CI: 2.7%-9.3%) based on diagnosis using commercial kits of the competitive enzyme-linked immunosorbent assay (CELISA) for Brucella abortus antibodies. Mean within-herd prevalence was found to be 25.9% (95% CI: 9.7%-53.1%) and brucellosis prevalence in an infected herd ranged from 9.1% to 50%. A risk factor could not be identified at the animal level but two risk factors were identified at the herd level: large herd size and history of abortion. The mean number of milking cows in a free-grazing herd (5.0) was significantly larger than a herd with a movement restricted (1.7, p < 0.001). CONCLUSIONS: Vaccination should be targeted at commercial large-scale farms with free-grazing farming to control brucellosis in cattle in and around Kampala city.

Spatial epidemiology of hospital-diagnosed brucellosis in Kampala, Uganda.

BACKGROUND: A retrospective case-control study was undertaken to examine the spatial risk factors for human brucellosis in Kampala, Uganda. METHODS: Information on age, sex and month of diagnosis was derived from records from plate agglutination tests undertaken at Mulago Hospital, Kampala. Information on Parishes (LC2s) where patients reside was sourced from the outpatient registration book. In-patient fracture cases were selected for use as controls using 1:1 matching based on the age, sex and month of diagnosis. The locations of cases and controls were obtained by calculating Cartesian coordinates of the centroids of Parish level (LC2) polygons and a spatial scan statistic was applied to test for disease clustering. Parishes were classified according to the level of urbanization as urban, peri-urban or rural. RESULTS: Significantly more females than males were found to show sero-positivity for brucellosis when compared with the sex ratio of total outpatients, in addition female brucellosis patients were found to be significantly older than the male patients. Spatial clustering of brucellosis cases was observed including around Mulago Hospital (radius = 6.8 km, p = 0.001). The influence of proximity to the hospital that was observed for brucellosis cases was not significantly different from that observed in the controls. The disease cluster was confounded by the different catchment areas between cases and controls. The level of urbanization was not associated with the incidence of brucellosis but living in a slum area was a significant risk factor among urban dwellers (odds ratio 1.97, 95% CI: 1.10-3.61). CONCLUSIONS: Being female was observed to be a risk factor for brucellosis sero-positvity and among urban dwellers, living in slum areas was also a risk factor although the overall risk was not different among urban, peri-urban and rural areas of the Kampala economic zone.

Serosurvey of Avian Influenza Viruses (H5, H7, and H9) and Associated Risk Factors in Backyard Poultry Flocks of Lahore District, Pakistan.

Rural poultry constitutes 56% of the total poultry population in Pakistan; however, epidemiological information about avian influenza viruses (AIVs) in backyard poultry flocks is lacking. A cross-sectional survey of villages of Lahore district was conducted from July 2009 to August 2009 using two-stage cluster sampling and probability proportional to size (PPS) sampling to estimate seroprevalence and its associated risk factors. A random selection of 35 clusters from 308 villages of Lahore were considered, and from each cluster, six chickens aged >2 months were selected. A total of 210 serum samples were collected and examined by the hemagglutination inhibition (HI) test for specific antibodies against AIV subtypes H5, H7, and H9. Overall weighted seroprevalence for AIVs was 65.2% (95% CI: 55.6-74.8%), and for subtype H5, H7 & H9 was 6.9% (95% CI: 10.8-23.0%), 0% (95% CI: 0-1.7%), and 62.0% (95% CI: 52.2-71.8%) respectively. However, none of the samples were positive for H7. The average flock size was 17.3 birds, and the main purpose of keeping poultry was for eggs/meat (70.6%, 95% CI: 59.7-81.4). A majority of them were reared in a semi-caged system (83%, 95% CI: 74.5-91.3). Backyard birds were received from different sources, that is, purchased from the market or received as a gift from friends or any NGO, and were 5.7 times more likely to become avian influenza (AI) seropositive than those that were not exposed to these sources (CI 95%: 2.0-716.0). Backyard birds which were received from different sources, that is, purchased from the market or received from friends or any NGO, were 5.7 times more likely to become AI seropositive compared to those that were not (CI 95%: 2.5-18.7). To reduce the risk of AIV in Pakistan, continuous surveillance of backyard poultry would be needed.

Coxiella burnetii abortion in a dairy farm selling artisanal cheese directly to consumers and review of Q fever as a bovine abortifacient in South America and a human milk-borne disease.

Coxiella burnetii is a highly transmissible intracellular bacterium with a low infective dose that causes Q fever (coxiellosis), a notifiable zoonotic disease distributed worldwide. Livestock are the main source of C. burnetii transmission to humans, which occurs mostly through the aerogenous route. Although C. burnetii is a major abortifacient in small ruminants, it is less frequently diagnosed in aborting cattle. We report a case of C. burnetii abortion in a lactating Holstein cow from a dairy farm producing and selling artisanal cheese directly to consumers in Uruguay, and review the literature on coxiellosis as a bovine abortifacient in South America and as a milk-borne disease. The aborted cow had severe necrotizing placentitis with abundant intratrophoblastic and intralesional C. burnetii confirmed by immunohistochemistry and PCR. After primo-infection in cattle, C. burnetii remains latent in the lymph nodes and mammary glands, with milk being a significant and persistent excretion route. Viable C. burnetii has been found in unpasteurized milk and cheeses after several months of maturing. The risk of coxiellosis after the consumption of unpasteurized dairy products, including cheese, is not negligible. This report raises awareness on bovine coxiellosis as a potential food safety problem in on-farm raw cheese manufacturing and sales. The scant publications on abortive coxiellosis in cattle in South America suggest that the condition has probably gone underreported in all countries of this subcontinent except for Uruguay. Therefore, we also discuss the diagnostic criteria for laboratory-based confirmation of C. burnetii abortion in ruminants as a guideline for veterinary diagnosticians.

Molecular prevalence of Coxiella burnetii in bulk-tank milk from bovine dairy herds: Systematic review and meta-analysis.

Coxiella burnetii is an obligate intracellular zoonotic bacterium that causes Q fever. Ruminants, including cattle, are broadly known to be reservoirs for this bacterium. Since 2006, many research groups have evaluated the herd-level prevalence of C. burnetii in cattle by molecular techniques on composite milk samples. This study explored the global C. burnetii herd-level prevalence from studies done on bovine bulk-tank milk (BTM) samples using PCR-based analysis. Also, moderators were investigated to identify sources of heterogeneity. Databases (CAB Abstracts, Medline via Ovid, PubMed, Web of Science and Google Scholar) were searched for index articles on C. burnetii prevalence in BTM samples by PCR published between January-1973 and November-2018. Numerous studies (1054) were initially identified, from which seventeen original publications were included in the meta-analysis based on the pre-defined selection criteria. These studies comprised 4031 BTM samples from twelve countries. A random-effects model was used because of considerable heterogeneity (I 2 = 98%) to estimate the herd-level prevalence of C. burnetii as 37.0%(CI95%25.2-49.5%). The average herd size appeared to account for a high level of the heterogeneity. No other moderators (geographic location, gross national income or notification criteria for Q fever) seemed to be determinant. This systematic evaluation demonstrated a high molecular prevalence of C. burnetii in BTM samples both in European and non-European countries, evidencing a widespread herd-level circulation of this agent in bovine dairy farms around the world. Meta-regression showed herd size as the most relevant moderator with the odds of a BTM sample testing positive doubling with every unit increase.

Serological Evidence of Human Infection with Coxiella burnetii after Occupational Exposure to Aborting Cattle.

Cattle are broadly deemed a source of Coxiella burnetii; however, evidence reinforcing their role in human infection is scarce. Most published human Q fever outbreaks relate to exposure to small ruminants, notably goats. Anti-phase II C. burnetii IgG and IgM were measured by indirect fluorescent antibody tests in 27 farm and veterinary diagnostic laboratory workers to ascertain whether occupational exposure to cattle aborting due to C. burnetii was the probable source of exposure. Four serological profiles were identified on the basis of anti-phase II IgG and IgM titres. Profile 1, characterised by high IgM levels and concurrent, lower IgG titres (3/27; 11.1%); Profile 2, with both isotypes with IgG titres higher than IgM (2/27; 7.4%); Profile 3 with only IgG phase II (5/27; 18.5%); and Profile 4, in which neither IgM nor IgG were detected (17/27; 63.0%). Profiles 1 and 2 are suggestive of recent C. burnetii exposure, most likely 2.5-4.5 months before testing and, hence, during the window of exposure to the bovine abortions. Profile 3 suggested C. burnetii exposure that most likely predated the window of exposure to aborting cattle, while Profile 4 represented seronegative individuals and, hence, likely uninfected. This study formally linked human Q fever to exposure to C. burnetii infected cattle as a specific occupational hazard for farm and laboratory workers handling bovine aborted material.

Farmer estimation of live bodyweight of cattle: implications for veterinary drug dosing in East Africa.

The ability of smallholder farmers and animal health workers to estimate live bodyweight can critically affect the likelihood of under- or over-dosing of veterinary compounds in decentralised systems where farmers administer a significant proportion of the veterinary treatments. A survey of 324 cattle owned by 170 farmers was conducted in Busia District, Kenya. Cattle were weighed on a standard calibrated scale and owners were asked to estimate the live weight of their animals. Weights were also estimated by a weigh-band and by local animal health workers. Cattle owners consistently underestimated the weights of their cattle; 85.7% of the cattle had their bodyweights underestimated by an average of 46.9% of their true weight. Furthermore, very few cattle (19.0%) had their weights estimated accurately to within +/-20% of their true weight by these farmers, an accuracy range important for drug dosing. This finding raises concerns of widespread under-dosing of cattle with trypanocidal and other veterinary drugs. Animal health workers were better at estimating live bodyweight of cattle; 76.6% of cattle were estimated accurately to within +/-20% of their true weight. It is possible to improve farmers' and animal health workers' ability to estimate accurately live bodyweight of cattle with appropriate training. Evidence of this was provided by animal health workers whose estimates improved over time as they received feedback of the true weights of different sizes of cattle from the standard scale.

Evaluating the effectiveness of trematocides against Fasciola gigantica and amphistomes infections in cattle, using faecal egg count reduction tests in Iringa Rural and Arumeru Districts, Tanzania.

BACKGROUND: Fasciolosis, caused by the liver fluke Fasciola gigantica, and paramphistomosis are widespread in cattle in Tanzania, and the use of trematocides is encouraged by the Government livestock extension officers. However, reduced efficacy of oxyclozanide against Fasciola gigantica and amphistomes (rumen flukes), and albendazole against F. gigantica, has been reported in some regions. This study was conducted to assess the efficacy of different trematocides against F. gigantica and amphistome infections in cattle at Iringa Rural and Arumeru Districts. METHODS: Cattle found with concurrent infection of F. gigantica and amphistomes were randomly grouped into six experimental groups. One control group was left untreated while five treatment groups were treated with one of five trematocides that include: albendazole, nitroxynil, oxyclozanide, closantel and triclabendazole. Post-treatment faecal sample collection was done on the day of treatment and again at 7, 14 and 28 days, from each cattle. The samples were processed by Flukefinder® method to recover and identify eggs. Assessment of the efficacy of the trematocides against F. gigantica and amphistomes was conducted using faecal egg count reduction (FECR) tests. RESULTS: The findings of the present study in both districts indicate that nitroxynil, oxyclozanide, closantel and triclabendazole are effective against patent F. gigantica infection, as the calculated FECR% for each trematocide was 100% by day 14 post-treatment. However, albendazole found to have reduced efficacy of against F. gigantica, as FECR% was 49% in Arumeru District and 89% in Iringa Rural District by day 14 post-treatment. Oxyclozanide was the only trematocide found to be effective against amphistomes with FECR of 99%. CONCLUSIONS: Albendazole had reduced efficacy against F. gigantica in cattle in Arumeru and Iringa Rural Districts, Tanzania. The reduced efficacy was prominent in Arumeru, where cattle are commonly treated with anthelmintics, than in Iringa Rural, where cattle are seldom treated.

Risk Factors for Avian Influenza H9 Infection of Chickens in Live Bird Retail Stalls of Lahore District, Pakistan 2009-2010.

This study was conducted to identify risk factors associated with AIV infections in live bird retail stalls (LBRS) in Lahore District, Pakistan. A cross-sectional survey of LBRS was conducted from December 2009-February 2010 using two-stage cluster sampling based on probability proportional to size. A total of 280 oropharyngeal swab sample pools were collected from 1400 birds in 8 clusters and tested by qRT-PCR for the matrix (M) gene of type A influenza virus and HA gene subtypes H9, H5 and H7. Thirty-four (34) samples were positive for the M gene, of which 28 were also positive for H9. No sample was found positive for H5 or H7. Data for 36 potential risk factors, collected by questionnaire, were analyzed by survey-weighted logistic regression and prevalence odds ratios (OR) for associated risk factors were calculated. A final multivariable model identified three risk factors for H9 infection in LRBS, namely obtaining birds from mixed sources (OR 2.28, CI95%: 1.4-3.7), keeping birds outside cages (OR 3.10, CI95%: 1.4-7.0) and keeping chicken breeds other than broilers (OR 6.27, CI95%: 1.7-23.2). Sourcing birds from dealers/wholesalers, keeping birds inside cages and avoiding mixing different breeds in cages could reduce the risk of H9 infections in LRBS.

Crisis, what crisis? Control of Rhodesian sleeping sickness.

There is an urgent need for cost-effective strategies for the sustainable control of Trypanosoma brucei rhodesiense (Rhodesian) sleeping sickness, which is a fatal zoonotic disease that has caused devastating epidemics during the past century. Sleeping sickness continues to be controlled by crisis management, using active case detection, treatment and vector control - activities that occur only during major epidemics; during the intervening periods, farmers and communities must fend for themselves. There are several methods for assessing the burden of this disease and there is a series of farmer-led methodologies that can be applied to reduce the burden of human and animal trypanosomiases.

Cross-sectional study of Fasciola gigantica and other trematode infections of cattle in Edu Local Government Area, Kwara State, north-central Nigeria.

BACKGROUND: Trematode infections of livestock are of global veterinary and public health importance causing serious economic losses. Majority of data on burden of trematode infections in Nigeria are based on abattoir surveys and there are very few data on herd level risk factors. The present study investigated the prevalence of, and herd level risk factors for, fasciolosis and other trematode infections in cattle in Edu Local Government Area (LGA). METHODS: A cross-sectional survey used two-stage study design to investigate cattle belonging to 65 households. Two questionnaires were administered for household-level and individual cattle-level data. Faecal and blood samples were obtained from the cattle. Logistic regression analyses were performed to determine risk factors for infections. RESULTS: Of 686 faecal samples analysed, 74.9 %, 16.1 %, 7.3 % and 1.2 % were positive for infections with Fasciola gigantica, paramphistomes, Dicrocoelium hospes and Schistosoma bovis respectively. Fasciola gigantica had higher prevalence in adult cattle (77.3 %) than weaners (62.5 %). Majority of co-infections was a combination of F. gigantica with paramphistomes 84/130 (64.6 %). Most (58.9 %) of the cattle belonged to FAMACHA© score 2. The mean packed cell volume (PCV) was 34.4 %. The sensitivity and specificity of FAMACHA© for anaemia (PCV < 24 %) were 18.2 and 96.9 %, respectively. Positive correlation was obtained between faecal egg counts for F. gigantica and paramphistomes (R = 0.15, P = 0.0001). Adult cattle were more likely to be infected with F. gigantica (odds ratio, OR: 1.94; Confidence Interval, CI: 1.19-3.16) than weaners. Cattle belonging to household heads aged between 40-59 years were more likely infected with paramphistomes (OR: 1.95; CI: 1.02-3.74) than those belonging to other age groups. Cattles from herds with size ≥ 100 were more likely infected with D. hospes than those from smaller herds (OR: 6.98; CI: 2.94-16.6). CONCLUSION: This study revealed high prevalence of infection with F. gigantica in Kwara State. The co-infections by F. gigantica and paramphistomes with a positive correlation should be considered during anthelmintic therapy. There is a need to optimise and validate the FAMACHA© for use in cattle based on breeds and variation in colour of ocular mucous membrane. Risk factors identified could assist in tailoring control strategies for various trematode infections to particular groups of farmers and cattle.

Ecological Monitoring and Health Research in Luambe National Park, Zambia: Generation of Baseline Data Layers.

Classifying, describing and understanding the natural environment is an important element of studies of human, animal and ecosystem health, and baseline ecological data are commonly lacking in remote environments of the world. Human African trypanosomiasis is an important constraint on human well-being in sub-Saharan Africa, and spillover transmission occurs from the reservoir community of wild mammals. Here we use robust and repeatable methodology to generate baseline datasets on vegetation and mammal density to investigate the ecology of warthogs (Phacochoerus africanus) in the remote Luambe National Park in Zambia, in order to further our understanding of their interactions with tsetse (Glossina spp.) vectors of trypanosomiasis. Fuzzy set theory is used to produce an accurate landcover classification, and distance sampling techniques are applied to obtain species and habitat level density estimates for the most abundant wild mammals. The density of warthog burrows is also estimated and their spatial distribution mapped. The datasets generated provide an accurate baseline to further ecological and epidemiological understanding of disease systems such as trypanosomiasis. This study provides a reliable framework for ecological monitoring of wild mammal densities and vegetation composition in remote, relatively inaccessible environments.

Movement Behaviour of Traditionally Managed Cattle in the Eastern Province of Zambia Captured Using Two-Dimensional Motion Sensors.

Two-dimensional motion sensors use electronic accelerometers to record the lying, standing and walking activity of cattle. Movement behaviour data collected automatically using these sensors over prolonged periods of time could be of use to stakeholders making management and disease control decisions in rural sub-Saharan Africa leading to potential improvements in animal health and production. Motion sensors were used in this study with the aim of monitoring and quantifying the movement behaviour of traditionally managed Angoni cattle in Petauke District in the Eastern Province of Zambia. This study was designed to assess whether motion sensors were suitable for use on traditionally managed cattle in two veterinary camps in Petauke District in the Eastern Province of Zambia. In each veterinary camp, twenty cattle were selected for study. Each animal had a motion sensor placed on its hind leg to continuously measure and record its movement behaviour over a two week period. Analysing the sensor data using principal components analysis (PCA) revealed that the majority of variability in behaviour among studied cattle could be attributed to their behaviour at night and in the morning. The behaviour at night was markedly different between veterinary camps; while differences in the morning appeared to reflect varying behaviour across all animals. The study results validate the use of such motion sensors in the chosen setting and highlight the importance of appropriate data summarisation techniques to adequately describe and compare animal movement behaviours if association to other factors, such as location, breed or health status are to be assessed.