Diagnosis of cattle diseases endemic to sub-Saharan Africa : evaluating a low cost decision support tool in use by veterinary personnel

Background: Diagnosis is key to control and prevention of livestock diseases. In areas of sub-Saharan Africa where private practitioners rarely replace Government veterinary services reduced in effectiveness by structural adjustment programmes, those who remain lack resources for diagnosis and might benefit from decision support. Methodology/Principal Findings: We evaluated whether a low-cost diagnostic decision support tool would lead to changes in clinical diagnostic practice by fifteen veterinary and animal health officers undertaking primary animal healthcare in Uganda. The eight diseases covered by the tool included 98% of all bovine diagnoses made before or after its introduction. It may therefore inform proportional morbidity in the area; breed, age and geographic location effects were consistent with current epidemiological understanding. Trypanosomosis, theileriosis, anaplasmosis, and parasitic gastroenteritis were the most common conditions among 713 bovine clinical cases diagnosed prior to introduction of the tool. Thereafter, in 747 bovine clinical cases estimated proportional morbidity of fasciolosis doubled, while theileriosis and parasitic gastroenteritis were diagnosed less commonly and the average number of clinical signs increased from 3.5 to 4.9 per case, with 28% of cases reporting six or more signs compared to 3% beforehand. Anaemia/pallor, weakness and staring coat contributed most to this increase, approximately doubling in number and were recorded in over half of all cases. Finally, although lack of a gold standard hindered objective assessment of whether the tool improved the reliability of diagnosis, informative concordance and misclassification matrices yielded useful insights into its role in the diagnostic process. Conclusions/Significance: The diagnostic decision support tool covered the majority of diagnoses made before or after its introduction, leading to a significant increase in the number of clinical signs recorded, suggesting this as a key beneficial consequence of its use. It may also inform approximate proportional morbidity and represent a useful epidemiological tool in poorly resourced areas

Occurrence of concurrent trypanosomosis, theileriosis, anaplasmosis and helminthosis in Friesian, Zebu and Sahiwal cattle in Uganda

An epidemiological investigation was conducted on farms in Tororo and Soroti districts of Uganda from January to February 2000 to determine the cause of reported persistent mortality of cattle. Blood and faecal material of 98 cattle comprising of 33 Friesians, 58 Zebu and 7 Sahiwal were examined. Results revealed that seven (7.1%) cattle had trypanosome infection, mainly due to Trypanosoma vivax and T. brucei, 17 (17.3%) Fasciola infection, 28 (28.6%) gastrointestinal nematode infection, 33 (33.7%) Theileria sp. infection and 13 (13.3%) Anaplasma marginale infection. Mixed infections were detected in 30%, 20.6% and 43% of the Friesian, Zebu and Sahiwal cattle respectively. Anaemia (PCV<25) was detected in 24%, 19% and 14% of the Friesian, Zebu and Sahiwal cattle respectively. Persistent mortality of cattle on these farms could have been due to either single or mixed parasitic infections probably exacerbated by malnutrition.The articles have been scanned in colour with a HP Scanjet 5590; 600dpi. Adobe Acrobat v.9 was used to OCR the text and also for the merging and conversion to the final presentation PDF-format.mn201

Development of Loop-Mediated Isothermal Amplification (LAMP) Assays for Rapid Detection of Ehrlichia ruminantium

<p>Abstract</p> <p>Background</p> <p>The rickettsial bacterium <it>Ehrlichia ruminantium </it>is the causative agent of heartwater, a potential zoonotic disease of ruminants transmitted by ticks of the genus <it>Amblyomma</it>. The disease is distributed in nearly all of sub-Saharan Africa and some islands of the Caribbean, from where it threatens the American mainland. This report describes the development of two different loop-mediated isothermal amplification (LAMP) assays for sensitive and specific detection of <it>E. ruminantium</it>.</p> <p>Results</p> <p>Two sets of LAMP primers were designed from the pCS20 and <it>sodB </it>genes. The detection limits for each assay were 10 copies for pCS20 and 5 copies for <it>sodB</it>, which is at least 10 times higher than that of the conventional pCS20 PCR assay. DNA amplification was completed within 60 min. The assays detected 16 different isolates of <it>E. ruminantium </it>from geographically distinct countries as well as two attenuated vaccine isolates. No cross-reaction was observed with genetically related Rickettsiales, including zoonotic <it>Ehrlichia </it>species from the USA. LAMP detected more positive samples than conventional PCR but less than real-time PCR, when tested with field samples collected in sub-Saharan countries.</p> <p>Conclusions</p> <p>Due to its simplicity and specificity, LAMP has the potential for use in resource-poor settings and also for active screening of <it>E. ruminantium</it> in both heartwater-endemic areas and regions that are at risk of contracting the disease.</p

Large-scale international validation of an indirect ELISA based on recombinant nucleocapsid protein of rift valley fever virus for the detection of IgG antibody in domestic ruminants

Diagnostic performance of an indirect enzyme-linked immunosorbent assay (I-ELISA) based on a recombinant nucleocapsid protein (rNP) of the Rift Valley fever virus (RVFV) was validated for the detection of the IgG antibody in sheep (n = 3367), goat (n = 2632), and cattle (n = 3819) sera. Validation data sets were dichotomized according to the results of a virus neutralization test in sera obtained from RVF-endemic (Burkina Faso, Democratic Republic of Congo, Mozambique, Senegal, Uganda, and Yemen) and RVF-free countries (France, Poland, and the USA). Cut-off values were defined using the two-graph receiver operating characteristic analysis. Estimates of the diagnostic specificity of the RVFV rNP I-ELISA in animals from RVF-endemic countries ranged from 98.6% (cattle) to 99.5% (sheep) while in those originating from RVF-free countries, they ranged from 97.7% (sheep) to 98.1% (goats). Estimates of the diagnostic sensitivity in ruminants from RVF-endemic countries ranged from 90.7% (cattle) to 100% (goats). The results of this large-scale international validation study demonstrate the high diagnostic accuracy of the RVFV rNP I-ELISA. Standard incubation and inactivation procedures evaluated did not have an adverse effect on the detectable levels of the anti-RVFV IgG in ruminant sera and thus, together with recombinant antigen-based I-ELISA, provide a simple, safe, and robust diagnostic platform that can be automated and carried out outside expensive bio-containment facilities. These advantages are particularly important for less-resourced countries where there is a need to accelerate and improve RVF surveillance and research on epidemiology as well as to advance disease control measures.The International Atomic Energy Agencyhttp://www.mdpi.com/journal/virusespm2021Medical Virolog

Multi-locus sequence typing of Ehrlichia ruminantium strains from geographically diverse origins and collected in Amblyomma variegatum from Uganda

Background: The rickettsial bacterium Ehrlichia ruminantium is the causative agent of heartwater in ruminants. A better understanding of the population genetics of its different strains is, however, needed for the development of novel diagnostic tools, therapeutics and prevention strategies. Specifically, the development of effective vaccination policies relies on the proper genotyping and characterisation of field isolates. Although multi-locus sequence typing (MLST) has been recently developed, only strains from geographically restricted collections have been analysed so far. The expansion of the MLST database to include global strains with different geographic origins is therefore essential. In this study, we used a panel of reference strains from geographically diverse origins and field samples of E. ruminantium detected from its vector, Amblyomma variegatum, in heartwater-endemic areas in Uganda. Results: A total of 31 novel alleles (six, four, six, three, two, five, three, and two for gltA, groEL, lepA, lipA, lipB, secY, sodB, and sucA loci, respectively) and 19 novel sequence types (STs) were identified. Both neighbour-joining and minimum spanning tree analyses indicated a high degree of genetic heterogeneity among these strains. No association was observed between genotypes and geographic origins, except for four STs from West African countries. When we performed six different tests for recombination (GeneConv, Bootscan, MaxChi, Chimaera, SiScan, and 3Seq) on concatenated sequences, four possible recombination events were identified in six different STs. All the recombination breakpoints were located near gene borders, indicating the occurrence of intergenic recombination. All four STs that localized to a distinct group in clustering analysis showed evidence of identical recombination events, suggesting that recombination may play a significant role in the diversification of E. ruminantium. Conclusions: The compilation of MLST data set across the African continent will be particularly valuable for the understanding of the existing genetic diversity of field isolates in African countries. Comprehensive information on the degree of cross-protection between strains and further understanding of possible relationships between genotypes and phenotypes such as vaccine efficacy are expected to lead to the development of region-specific vaccination strategies

Concordance matrices (κ) of participants’ diagnoses and diagnoses calculated by the DST using clinical signs recorded during the study.

<p>κ-values for each DST diagnosis in columns are indicated for each participant diagnosis in rows. Main diagonal (mostly boldface numbers) indicating agreement between like diagnoses, other cells indicating possible cross-agreement between differing diagnoses. Boldface numbers indicate at least ‘fair’ agreement (κ ≥0.2); non-bold, positive numbers indicate slight agreement (0.2> κ >0); negative numbers indicate no cross-agreement, i.e. less than that expected by chance.</p><p>¹Other: any participant’s diagnosis other than the eight conditions listed on the DST; a DST diagnosis of ‘other’ resulted when none of its 16 signs was recorded for a case.</p>2<p>SCH: schistosomosis. [Further abbreviations as <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0040687#pone-0040687-t003" target="_blank">Table 3</a>.].</p

Approximate proportional morbidity in cattle in the eastern region of Uganda based on participating veterinary and animal health officers’ diagnoses and those suggested by the DST.

<p>Numbers of individual diagnoses made in each of the two phases of the study are expressed as a proportion of all diagnoses made in that phase. Phase-1 (n = 713 cases) was prior to, and Phase-2 (n = 747 cases) after, the introduction of the diagnostic decision support tool.</p

Misclassification matrices, (a) M<i>_v</i> and (b) M<i>_c</i>, of participants’ diagnoses and diagnoses calculated by the DST using clinical signs recorded during the study.

<p>In (a), <b>M</b><b><i>_v</i></b>, proportions of each participant diagnosis are shown for each DST diagnosis in columns summating to 1. In (b), <b>M</b><b><i>_c</i></b>, proportions of each DST diagnosis are shown for each participant diagnosis in columns summating to 1.</p><p>Leading diagonal indicating agreement, other cells indicating disagreement.</p><p>¹NA: not applicable. [Other abbreviations as <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0040687#pone-0040687-t004" target="_blank">Table 4</a>.].</p

Frequency distribution of numbers of clinical signs recorded for bovine cases by Ugandan veterinary and animal health officers during the two phases of the study.

<p>Histogram showing the frequency distribution of number of clinical signs per case observed during each of the two phases of the study, prior to and after the introduction of the diagnostic decision support tool.</p

Template casebook used by participating Ugandan veterinary and animal health officers to record data in Phase-2 of the study.

<p>The template casebook was provided in the form of a printed notebook with the decision support tool appearing on each left hand page in landscape orientation so that clinical signs observed could be marked directly on it.</p