Dickson Stuart Tayebwa

Genetic mutations in sodium channel domain II and carboxylesterase genes associated with phenotypic resistance against synthetic pyrethroids by Rhipicephalus (Boophilus) decoloratus ticks in Uganda

We previously reported emergence of super synthetic pyrethroid (SP) resistant Rhipicephalus (Boophilus) decoloratus ticks in Uganda. This study investigated the genetic basis of phenotypic resistance against SP in R. (B.) decoloratus and sought to identify novel diagnostic mutations for rapid detection of SP resistance in the above tick species. Genomic DNA was extracted from pooled larvae of 20 tick populations (19 of known SP susceptibility and 1 unknown susceptibility). The voltage sensitive sodium channel (VSSC) domain II S4-5 linker (SP target) and partial carboxylesterase (SP metabolizing enzyme) genes were amplified by PCR, cloned and sequenced. The resultant sequences were analyzed to determine single nucleotide polymorphisms (SNPs) associated with phenotypic resistance in the various tick populations investigated. Novel SNPs that introduced Eco RI and Eco RII restriction sites in carboxylesterase gene were identified in silco and validated with restriction fragment length polymorphism (RFLP) against 18 tick populations of known SP susceptibility. The study identified a super knock down resistance (kdr) mutation T58C in R. (B.) decoloratus VSSC associated with stable SP resistance. We further identified multiple nonsynonymous mutations in carboxylesterase of SP resistant ticks; one of which conferred novel EcoRII (G195C) restriction site for PCR-RFLP detection of SP resistance. In conclusion, this study is the first to report super kdr mutation in sodium channel domain II and multiple mutations in carboxylesterase genes that may concurrently mediate stable resistance against synthetic pyrethroids in R. (B.) decoloratus ticks from Uganda. The Eco RII based PCR-RFLP is a useful tool for rapid detection of stable SP resistant R. (B.) decoloratus ticks.

Retrospective study on cattle and poultry diseases in Uganda

Abstract Cattle and poultry enterprises are among the major contributors to food security and socioeconomic empowerment of households in Uganda. However, various diseases constrain their productivity. A two-year retrospective study between April 2012 and March 2014 was conducted using records for cattle and poultry diseases diagnosed at the Central Diagnostic Laboratory (CDL) to determine prevalent diseases in Uganda. The laboratory received 836 samples from poultry (36.3%) and cattle (63.7%). Of the 836 samples, 47.5% had a definitive diagnosis of disease causation. Most of the cattle and poultry diseases diagnosed were protozoan diseases (39.3%) followed by bacterial (21.4%), viral (17.1%), helminthiasis (11.1%), nutritional diseases (4%) and others (7.1%). For poultry, viral diseases (29.5%) and protozoan diseases (27.1%) especially newcastle disease (44.3%) and coccidiosis (100%) respectively, were the most diagnosed. While for cattle, hemo-protozoan parasites (52.1%) were the most prevalent, of which 92.9% were east coast fever infection. Bacterial infection (20.5%) in cattle were the second most diagnosed diseases and mastitis was the most diagnosed (46.2%). In summary, coccidioisis, collibacillosis, newcastle disease, gumboro disease, and avian helminthiasis were the most prevalent poultry diseases while in cattle, east coast fever, helminthiasis, mastitis, brucellosis and rabies were the most frequently diagnosed diseases. This study has identified the major diseases that hinder poultry and cattle production in Uganda. The data generated by CDL could be used for surveillance, monitoring and designing strategic interventions for control of poultry and cattle diseases in Uganda.

Prevalence of endometritis and its associated risk factors in dairy cattle of Central Uganda.

Aims: Endometritis is a major postpartum disease that affects dairy cattle productivity which is accompanied by heavy economic losses to the farmer. The status of Clinical endometritis (CE) and Sub-Clinical Endometritis (SCE) in sub-Saharan Africa is poorly understood, thus the study was carried out to provide information on the prevalence and associated risk factors that are responsible for the occurrence of SCE and CE in dairy cattle of Central Uganda. Study Design: This was a prospective study involving 140 dairy cattle within 60 days postpartum from 35 commercial dairy farms in Central Uganda. The dairy herds were visited in both the dry (n=90) and wet season (n=50) and subsequent visits were conducted after 3 months and 5 months to collect data for reproductive performance Original Research Article Tayebwa et al.; AJEA, 7(3): xxx-xxx, 2015; Article no.AJEA.2015.115 155 Methodology: A metricheck® device was used to collect the cervico-vaginal discharge which was examined for color, odor, texture, and measurement of its pH during the postpartum period for diagnosis of endometritis. Further examination of the reproductive tract was carried out using a vaginal speculum and subsequently rectal palpations were performed. The objectives of the study were to determine the prevalence of CE and SCE in the Central Uganda and assess the risk factors involved. Results: In this study, the prevalence of CE and SCE was established at 3.6% and 18.6% respectively, this burden was slightly higher in the wet than in the dry season but with no statistical significance P=0.126. Dairy cattle that had calved more than three times were shown to be associated with a higher body appearance (BCS > 3) than those that had calved down fewer times. Dystocia, Retained After Birth and Abortion were identified as associated risk factors (P = 0.00) to SCE and CE whereas infertility and Prolonged days calving to conception (>90 d) were postpartum implications (P=0.00) associated with SCE and CE in this study. This would be attributed to the poor management of postpartum dairy cattle in the farms visited as no farm was found with a maternity pen. SCE caused infertility in 65.5% of the dairy cattle whereas the CE is a major influencing factor to long calving to first AI and calving to conception interval (306d±90.6), On basis of reproduction, there was no major difference towards use of AI or Natural service. Conclusion: Management of endometritis in the region should address pre-partum and postpartum dairy herd management through improved extension service delivery and technical farm support to construct maternity pens, Endometritis is a multifactorial disease that requires a multidisciplinary approach to boost nutrition and health thus reducing incidence of risk factors such as dystocia and Retained fetal birth) .subsequent studies should be carried out to explore the national burden of SCE and ascertain the cause of the abortion.

C190A knockdown mutation in sodium channel domain II of pyrethroid-resistant Rhipicephalus appendiculatus

This study investigated the target site mutations in the partial sequence of voltage-sensitive sodium channel (VSSC) domain II of synthetic pyrethroid (SP)-resistant Rhipicephalus appendiculatus. Genomic DNA was extracted from seven tick populations (two susceptible and five resistant) collected from central, eastern and southwestern Uganda. The PCR amplicons of the VSSC domain II were cloned and sequenced to determine novel single nucleotide polymorphisms (SNP). A non-synonymous mutation C78 A corresponding to C190 A was found in all the five SP-resistant ticks. The C78 A mutation led to amino acid substitution from leucine to isoleucine (L21I) which was previously reported to confer knockdown (kdr) mutation in R. (Boophilus) microplus. The genetic confirmation of SP-resistant R. appendiculatus in central and southwestern Uganda calls for an urgent strategy for controlling the ticks.

The effects of nitidine chloride and camptothecin on the growth of Babesia and Theileria parasites

The treatment of bovine and equine piroplasmosis is limited to diminazene aceturate (DA) and imidocarb dipropionate. To address this challenge, we need to explore novel drug compounds and targets. Topoisomerases are potential drug targets because they play a vital role in solving topological errors of DNA strands during replication. This study documented the effectiveness of topoisomerase inhibitors, nitidine chloride (NC) and camptothecin (Cpt), on the growth of Babesia and Theileria parasites. The half maximal inhibitory concentrations (IC50s) against B. bovis, B. bigemina, B. caballi, and T. equi were 1.01 ± 0.2, 5.34 ± 1.0, 0.11 ± 0.03, and 2.05 ± 0.4 μM for NC and 11.67 ± 1.6, 4.00 ± 1.0, 2.07 ± 0.6, and 0.33 ± 0.02 μM for Cpt, respectively. The viability experiment revealed that 4, 10, and 4 μM treatments of NC or 48, 8, and 8 μM treatments of Cpt were sufficient to stop the in vitro regrowth of B. bovis, B. bigemina, and B. caballi, respectively. However, T. equi regrew in all of the concentrations used. Moreover, increasing the concentration of NC and Cpt to 16 μM and 1.2 μM (8 × IC50) did not eliminate T. equi. The micrographs of B. bigemina and B. caballi taken at 24 h and 72 h showed deformed merozoites and remnants of parasites within the red blood cell (RBC), respectively. The treatments of 25 mg/kg DA and 20 mg/kg NC administered intraperitoneally and 20 mg/kg NC given orally showed 93.7, 90.7, and 83.6% inhibition against Babesia microti (B. microti), respectively, compared to the untreated group on day 8. In summary, NC and Cpt were effective against Babesia and Theileria parasites in vitro. Moreover, 20 mg/kg NC administered intraperitoneally was as effective as 25 mg/kg DA against B. microti in mice and showed no toxic symptoms in mice. The results indicate that NC may, after further evaluations, prove to be an alternative drug against bovine and equine piroplasmoses.

Food Safety Analysis of Milk and Beef in Southwestern Uganda

Background Inorganic pollutants in milk and beef are of major public health concern; however, information in Africa is still limited due to low food safety monitoring practices. In this study, we established levels of lead (Pb), zinc (Zn), cadmium (Cd), copper (Cu), and iron (Fe) in milk and beef and obtained the estimated daily intake (EDI) and incremental lifetime cancer risk (ILCR) as measures of risk to the Ugandan population. Materials and Methods This was a cross-sectional study in which a total of 40 samples of milk and beef were collected from Bushenyi district in southwestern Uganda. Samples were analyzed by atomic absorbance spectrophotometer, and the EDI and ILCR were computed using the US EPA reference values. Results and Discussion Heavy metal concentrations were highest in the order of Zn > Fe > Pb > Cu in milk samples, while in beef samples, concentrations were highest in the order of Zn > Pb > Fe > Cu and no Cd was detected. Furthermore, beef had significantly higher (P < 0.05) Pb and Fe concentrations than milk. The EDI was highest in children, and this was followed by very high ILCR levels, showing that milk and beef are not safe for children in Uganda. Bearing in mind that a high HI was shown, beef and milk from these regions are not recommended for consumption especially by children although more studies remain to be conducted. Conclusion Heavy metals in milk and beef of Uganda may predispose the indigenous community to cancer and other health-related illnesses, showing a need for improved food safety screening to promote food safety.