Assessment of genotyping array performance for genome-wide association studies and imputation in African cattle.

BACKGROUND: In cattle, genome-wide association studies (GWAS) have largely focused on European or Asian breeds, using genotyping arrays that were primarily designed for European cattle. Because there is growing interest in performing GWAS in African breeds, we have assessed the performance of 23 commercial bovine genotyping arrays for capturing the diversity across African breeds and performing imputation. We used 409 whole-genome sequences (WGS) spanning global cattle breeds, and a real cohort of 2481 individuals (including African breeds) that were genotyped with the Illumina high-density (HD) array and the GeneSeek bovine 50 k array. RESULTS: We found that commercially available arrays were not effective in capturing variants that segregate among African indicine animals. Only 6% of these variants in high linkage disequilibrium (LD) (r2 > 0.8) were on the best performing arrays, which contrasts with the 17% and 25% in African and European taurine cattle, respectively. However, imputation from available HD arrays can successfully capture most variants (accuracies up to 0.93), mainly when using a global, not continent-specific, reference panel, which partially reflects the unusually high levels of admixture on the continent. When considering functional variants, the GGPF250 array performed best for tagging WGS variants and imputation. Finally, we show that imputation from low-density arrays can perform almost as well as HD arrays, if a two-stage imputation approach is adopted, i.e. first imputing to HD and then to WGS, which can potentially reduce the costs of GWAS. CONCLUSIONS: Our results show that the choice of an array should be based on a balance between the objective of the study and the breed/population considered, with the HD and BOS1 arrays being the best choice for both taurine and indicine breeds when performing GWAS, and the GGPF250 being preferable for fine-mapping studies. Moreover, our results suggest that there is no advantage to using the indicus-specific arrays for indicus breeds, regardless of the objective. Finally, we show that using a reference panel that better represents global bovine diversity improves imputation accuracy, particularly for non-European taurine populations.

Efficacy of oral afoxolaner plus milbemycin oxime chewables against induced infestations with Dermacentor reticulatus in dogs.

The efficacy of afoxolaner plus milbemycin oxime (AFX + MO) combination chewables (NexGard Spectra®, Merial) and AFX single-entity chewables (NexGard®, Merial) against induced infestations with Dermacentor reticulatus ticks was evaluated in dogs. Thirty dogs were assigned to blocks of three animals each based on pre-allocation tick counts and were randomly allocated to one of three groups: untreated (control), treated with a combination of AFX + MO chewables to be as close as possible to the minimum effective dose of AFX + MO (2.5 + 0.5 mg per kg body weight), and treated with a combination of NexGard® chewables to be as close as possible to the minimum effective dose of AFX (2.5 mg per kg body weight). Treatments were administered orally once on day 0. Starting 2 days before treatment administration, each dog was infested with approximately 50 ticks weekly for six consecutive weeks. Live ticks were counted at ∼48 h post-treatment (removal count) and at ∼48 h (in situ counts) and ∼72 h (removal counts) following each post-treatment infestation. Treatment with both AFX + MO and NexGard® chewables rapidly eliminated the existing tick infestations (100 % efficacy) within 2 days following treatment administration. Weekly re-infestations were controlled for a minimum of 5 weeks with the efficacy ranging from 92.2 to 99.7 % based on ∼48 h post-treatment in situ counts and between 99.0 and 100 % based on ∼72 h post-treatment removal counts (p < 0.0001 at each occasion). This study demonstrated a high efficacy of both AFX + MO chewable and NexGard® chewable treatments against infestations of dogs with D. reticulatus ticks for at least 5 weeks. In addition, this study indicated no interference between the two compounds with respect to the acaricidal activity provided by AFX.

Efficacy of a single dose of milbemycin oxime/praziquantel combination tablets, Milpro(®), against adult Echinococcus multilocularis in dogs and both adult and immature E. multilocularis in young cats.

Two single-site, laboratory, negatively controlled, masked, randomised dose confirmation studies were performed: one in dogs, the other in cats. After a period of acclimatisation, both the dogs and cats were orally infected with Echinococcus multilocularis protoscoleces. In the dog study, 10 dogs received a single dose of Milpro® tablets at a minimum dose of 0.5 mg/kg milbemycin oxime and 5 mg/kg praziquantel 18 days post-infection and 10 dogs received no treatment. In the cat study, 10 cats received a single dose of Milpro® tablets at a minimum dose of 2 mg/kg milbemycin oxime and 5 mg/kg praziquantel 7 days post-infection, 10 cats received a single dose of the treatment 18 days post-infection and 10 cats remained untreated. In both studies, intestinal worm counts were performed 23 days post-infection at necropsy. No worms were retrieved from any of the 30 treated animals. Nine of 10 control dogs had multiple worms (geometric mean 91, arithmetic mean 304) and all 10 control cats had multiple worms (geometric mean 216, arithmetic mean 481). The difference in worm counts between all three treated groups and their controls was highly significant (ANOVA p values of log transformed data <0.0001). Efficacy of 100 % was demonstrated for the elimination of adult E. multilocularis in dogs and cats as well as for elimination of immature E. multilocularis in cats as evidenced by the effectiveness of treatment 7 days post-infection. The treatments were well accepted and tolerated, and there were no adverse drug reactions observed.

One-month comparative efficacy of three topical ectoparasiticides against adult brown dog ticks (Rhipicephalus sanguineus sensu lato) on mixed-bred dogs in controlled environment.

This study was designed to compare the therapeutic and residual efficacy for 1 month of three topical ectoparasiticides on mixed-bred dogs against the brown dog tick, Rhipicephalus sanguineus. Adult dogs (n = 32, 10.8-18.4 kg BW) were allocated to 4 groups (n = 8) and infested with 50 adult ticks on days -8, -2, 7, 14, 21, and 28. Within each group, dogs were treated topically on day 0 with a control solution (CS), Vectra 3D (DPP), Frontline Plus (FM), or K9 Advantix (IP). Ticks were enumerated on dogs 24 h after treatment and each subsequent tick infestation by in situ thumb count assessment without removal and at 48 h by combing and removal. Acaricidal efficacy was calculated using arithmetic means for all 24 and 48 h tick count assessments. From 42 to 56% of the total, infested ticks were found on dogs 48 h post-challenge in the CS group. Therapeutic efficacy for all treatments ranged from 45.5 to 64.6% after 48 h of infestation. Residual efficacy after FM treatment was consistently lower compared to DPP or IP treatments at the 24 h assessments on days 8, 22, 23, and 29. Residual efficacy measured at this last time point was 94.8% for DPP, 83.1% for IP, and 46.9% for FM. This study demonstrates that permethrin-based formulations (DPP and IP) provided a quicker onset of residual protection against brown dog ticks compared to FM. Although DPP and IP are both permethrin-based formulations, DPP exhibited consistently higher residual acaricidal efficacies and was the only treatment that provided >90% protection for 1 month at 24 h post challenge.

Expellency, anti-feeding and speed of kill of a dinotefuran-permethrin-pyriproxyfen spot-on (Vectra®3D) in dogs weekly challenged with adult fleas (Ctenocephalides felis) for 1 month-comparison to a spinosad tablet (Comfortis®).

This study was designed to compare the efficacy of two ectoparasiticides against adult fleas on dogs: a topical (DPP, dinotefuran-permethrin-pyriproxyfen) and a systemic (S, spinosad). Dogs (n = 48; 10.21-22.86 kg BW) were allocated to six groups of eight dogs each (C1, C4, DPP1, DPP4, S1, S4). Dogs in the treated groups were administered a topical (3.6 mL of DPP) or a tablet (665 or 1040 mg of S) on day 0. Infestations with 100 unfed fleas (Ctenocephalides felis) occurred on days -6, -1, 2, 7, 14, 21 and 28. An additional untreated group (QC, n = 6) was involved to evaluate the flea-anti-feeding efficacy. These dogs were infested once with 150 fleas prior to combing of at least 50 live fleas from each dog 5 or 10 min after infestation. In the treated group, dislodged dead and moribund fleas were collected from dogs 5, 10, 15 and 60 min (DPP1, S1) or 5, 10, 30 and 240 min (DPP4, S4) post-treatment and subsequent flea infestations on pans placed underneath the cages. Fleas were counted and removed from dogs by combing 1 (C1, DPP1, S1) or 4 h (C4, DPP4, S4) post-treatment and subsequent infestations. Quantitative PCR analysis of the canine cytochrome b gene was conducted on dislodged fleas collected from treated and control (QC) dogs 5 and 10 min after post-treatment infestations. The number of gene copies was used as a marker of blood volume ingested by fleas. Dislodgeability and insecticidal efficacy were calculated using arithmetic means. A rapid onset of killing was observed for DPP with 12.7 % of dead and moribund fleas being dislodged in average from dogs as soon as 5 min after infestation. DPP exhibited a significantly higher and sustained speed of kill than S. The average insecticidal efficacy was 86 ± 8.8 and 95.3 ± 2.1 % with DPP, whereas it was only 33.7 ± 19.9 and 57.6 ± 18.6 % with S at respectively 1 and 4 h after weekly reinfestations. The DPP combination significantly inhibited the feeding of fleas (89 % reduction) up to onset of flea mortality for 1-month post-treatment.

Comparative Speed of Kill, Repellent (anti-feeding) and Acaricidal Efficacy of an Imidacloprid/Flumethrin Collar (Seresto®) and a Fipronil/(S)-Methoprene/Eprinomectin/Praziquantel Spot-on (Broadline®) against Ixodes ricinus (Linné, 1758) on Cats.

Speed of kill, repellent (anti-feeding) and acaricidal efficacy of an imidacloprid 10 % (w/w) /flumethrin 4.5 % (w/w) collar (Seresto(®), Bayer) and a spot-on formulation of fipronil 8.3 % (w/v) /(S)-methoprene 10 % (w/v) /eprinomectin 0.4 % (w/v) /praziquantel 8.3 % (w/v) (Broadline(®), Merial) against artificiallyinduced infestations with Ixodes ricinus on cats, were assessed in a parallel group design, randomised, controlled study. Twenty-four cats were included and randomly allocated to treatment groups or a non-treated control group. Starting on Day (D) 7 after treatment until D28, cats were each infested with 50 I. ricinus at weekly intervals. Ticks were counted in situ on the cats at 6, 12 and 24 h and upon removal 48 h after each infestation. Based on arithmetic means, Seresto(®) proved to be 100 % effective against adult I. ricinus at all assessment times (6, 12, 24 and 48 h after infestation) throughout the month-long study. Broadline(®) was 0 % to 16.7 % effective at 6 h, 26.8 % to 50.0 % effective at 12 h, while at 24 h after infestation efficacy peaked at 81.5 % on D15 declining to 31.5 % on D29. Based on the 48 h tick counts, the efficacy of Broadline(®) peaked at 100 % on D16 after treatment and decreased to 83.2 % by D30. The Seresto(®) collar provided significantly faster speed of kill and better persistent acaricidal effectiveness against Ixodes ricinus on cats compared to Broadline(®) spot-on. The additional repellent (anti-feeding) effect of Seresto(®) prevents parasites from taking a blood meal and thereby reduces the risk of vector-borne disease pathogen transmission.

Chemical Compatibility and Safety of Imidacloprid/Flumethrin Collar (Seresto®) Concomitantly Used with Imidacloprid/Moxidectin (Advocate®, Advantage® Multi) and Emodepside/Praziquantel (Profender®) Spot-on Formulations.

Safety of concomitant use of veterinary products is of clinical interest. A series of studies was performed to evaluate the chemical compatibility and short term dermal and systemic safety of an imidacloprid/flumethrin collar (Seresto(®)/ Foresto(®), Bayer) used concomitantly with spot-on or tablet formulations.Chemical compatibility was evaluated in-vitro (study reference A) on collar pieces, followed by two small, non-controlled clinical studies (study reference B) in both, cats and dogs. The studies showed, that certain solvents affected the collar in-vitro, but not in their marketed formulations.Dermal and systemic safety of different spot-on or tablet formulations was first evaluated in a small, non-controlled clinical study (study reference C) in cats and dogs, via clinical observations only, followed by controlled clinical safety studies of concomitant use with imidacloprid/ moxidectin (Advocate(®)/ Advantage(®) Multi, Bayer) in dogs and cats (study reference D) and emodepside/ praziquantel (Profender(®), Bayer) in cats (study reference E), assessing safety aspects by clinical observations and statistical analyses of hematology and clinical chemistry parameters compared to baseline values and between treated and control groups.Dermal safety findings over all clinical studies (study references B to E) matched those already described for the respective products and included transient cosmetic changes (oily hair and crystal formation) at the site of spot-on application and broken hair, transient alopecia and skin alterations at the site of collar application. There were no indications of these findings aggravating under the conditions of concurrent use. There were no systemic safety findings of clinical significance in any of the clinical safety studies (study reference C to E). Assessment of blood parameters revealed some deviations from baseline levels and from the reference range in dogs as well as in cats, but no clinical relevance could be deduced. Hematology and clinical chemistry results confirmed the safety of the concomitant treatment. It is concluded that Seresto(®) is chemically compatible with solvents used in major spot-on formulations on the market and is dermally and systemically safe for adult dogs and cats when used concomitantly with Advocate(®) and Profender(®) spot-on formulations.

Phenotypic and genotypic characterization of acaricide resistance in Rhipicephalus microplus field isolates from South Africa and Brazil.

Rhipicephalus (Boophilus) microplus is one of the most successful ticks infesting cattle around the world. This highly-invasive species transmits cattle parasites that cause cattle fever leading to a high socio-economic burden. Tick eradication programs have often failed, due to the development of acaricide resistance. Here we characterize acaricide resistance in a large number of tick isolates from regions in South Africa (KwaZulu Natal, Mpumalanga, Western & Eastern Cape provinces) and two Brazilian regions. By means of Larval Packet Tests (LPT's) acaricide resistance was evaluated against five commonly used acaricides (chlorfenvinphos, fipronil, deltamethrin, amitraz, and ivermectin). Furthermore, the coding region containing the knock down resistance (kdr) mutation, known to result in pyrethroid resistance, was sequenced. Resistance to at least one acaricide class was reported in each of the five regions, and a high proportion of tick isolates exhibited multi-resistance to at least two acaricide classes (range: 22.2-80.0%). Furthermore, resistance ratios (RR) showed high spatial variation (intercontinental, as well as regional) but low regional spatial autocorrelation. Previous and current acaricide use correlated with current RR, and several combinations of acaricide RR were positively correlated. Moreover, fipronil resistance tended to be higher in farms with more intense acaricide use. The kdr-mutations provided the ticks a fitness advantage under the selection pressure of synthetic pyrethroids based on population (kdr-allele frequency) and individual level data (genotypes). The data show the threat of acaricide (multi-)resistance is high in Brazil and South Africa, but acaricide specific levels need to be assessed locally. For this purpose, gathering complementary molecular information on mutations that underlie resistance can reduce costs and expedite necessary actions. In an era of human-caused habitat alterations, implementing molecular data-driven programs becomes essential in overcoming tick-induced socio-economic losses.

Acaricide resistance status of livestock ticks from East and West Africa and in vivo efficacy of acaricides to control them.

Through a collaborative effort across six Sub-Saharan African countries, using recognized international assessment techniques, 23 stocks of three tick species (Rhipicephalus microplus, Rhipicephalus appendiculatus and Amblyomma variegatum) of economic importance for rural small holder farming communities from East and West Africa were collected from cattle, and evaluated in in vitro larval packet tests (LPT). The results demonstrated medium to high resistance to chlorfenvinphos and amitraz across species. Rhipicephalus microplus demonstrated high level alpha-cypermethrin and cypermethrin resistance. Stocks of A. variegatum (West Africa) and R. appendiculatus (Uganda) demonstrated medium level ivermectin resistance. The four least susceptible stocks (East and West African R. microplus, A. variegatum and R. appendiculatus) were taken into in vivo controlled cattle studies where fipronil was found effective against West and East African R. microplus isolates although persistent efficacy failed to reach 90%. Cymiazole and cypermethrin, and ivermectin based acaricides were partially effective against R. microplus without persistent efficacy. Flumethrin spray-on killed A. variegatum within 72 h for up to 10 days posttreatment, however product application was directly to tick attachment sites, which may be impractical under field conditions. A flumethrin pour-on formulation on goats provided persistent efficacy against A. variegatum for up to one-month. Therapeutic control was achieved against R. appendiculatus through weekly spraying cattle with flumethrin, amitraz or combined cymiazole and cypermethrin. A fipronil pour-on product offered four-week residual control against R. appendiculatus (with slow onset of action). Few studies have assessed and directly compared acaricidal activity in vitro and in vivo. There was some discordance between efficacy indicated by LPT and in vivo results. This observation calls for more research into accurate and affordable assessment methods for acaricide resistance. No single active or product was effective against all three tick species, emphasising the need for the development of alternative integrated tick management solutions.

Imidacloprid 10 % / flumethrin 4.5 % collars (Seresto®, Bayer) successfully prevent long-term transmission of Ehrlichia canis by infected Rhipicephalus sanguineus ticks to dogs.

The objective of this study was to determine the empirical efficacy of imidacloprid 10 %/flumethrin 4.5 % (Seresto®) collars in preventing long-term transmission of Ehrlichia canis by infected Rhipicephalus sanguineus ticks to dogs. The study was a parallel group design, single centre, randomised, non-blinded, controlled, long-term efficacy study. The treatment group of 8 dogs was fitted with Seresto® collars, the untreated control group of 8 dogs received no collars. Ehrlichia canis-infected ticks were released into the dogs sleeping quarters at 14-day intervals up to Day +378. Control group dogs infected with E. canis were continuously replaced to keep the control sample size constant, and a total of 39 control dogs were required. The final clinical examination and blood sampling occurred on Day +420. The primary assessment criterion was the number of dogs infected with E. canis, as confirmed by IFA and PCR, and the secondary criterion was the acaricidal efficacy based on tick counts. All scheduled blood samples taken were subject to analyses for both PCR and IFA, but only positive cases are discussed. Up to Day +378, none of the collar-treated dogs were infected with E. canis, whereas 34 of the 35 untreated dogs enrolled before Day +371 were infected. The acaricidal efficacy of the collar ranged from 90 % to 100 % for the duration of the assessment period.

Prophylactic treatment of flea-infested dogs with an imidacloprid / flumethrin collar (Seresto®, Bayer) to preempt infection with Dipylidium caninum.

The objective of the study was to determine the sustained effectiveness of 10 % imidacloprid and 4.5 % flumethrin, incorporated in a slow-release matrix collar, in preventing Dipylidium caninum infection in dogs after repeated laboratory infestations with fleas infected with metacestodes of this tapeworm. Efficacy against infection with D. caninum was evaluated by infesting 16 dogs with cat fleas (Ctenocephalides felis) on study days 7, 14, 21, 28, 35 and 42, from batches suitably infected with D. caninum metacestodes. Prior to each post-treatment infestation the D. caninum infection rate for the fleas was determined by microscopically examining 100 fleas for D. caninum metacestodes. The D. caninum prevalence in the fleas used for infestations ranged from 23 % to 52 %. Medicated collars were fitted to 8 of the dogs on study day 0. The weight of the IVP collars varied between 35.48 g and 38.48 g (average 37.16 g), whilst animal weight varied between 12.20 kg and 17.98 kg (treated group, n = 8, average 14.79 kg). Seven days later infestation of each of the 16 dogs with 250 fleas commenced. Infestations continued at weekly intervals until Day 42 with efficacy against fleas evaluated 24 hours after each infestation. From Days 21 to 74, infection of the dogs with D. caninum was verified (daily examination of faeces and cages for the presence of expelled proglottids). Calculation of prophylactic effectiveness of the collars in preventing infection with D. caninum was based on the difference in geometric mean numbers of scoleces between groups at necropsy on Day 75. Effective prevention of infection with D. caninum was found to be 96.6 %. Efficacy of the collars against fleas was ≥ 99.9 % for the duration of the assessment period. Newly acquired infestations of fleas are rapidly eliminated by the insecticidal components of the medicated collars over a period of several months. In the event of fleas being infected with metacestodes, with D. caninum can be prevented in collared dogs, concurrently reducing the likelihood of transmission to humans.

Efficacy of a novel fipronil spot-on for the treatment and control of induced infestations of adult cat fleas (Ctenocephalides felis) and castor bean ticks (Ixodes ricinus) on cats.

Cat fleas (Ctenocephalides felis) and the castor bean tick (Ixodes ricinus) cause discomfort and health effects due to bites and ingestion of blood and they serve as vectors for several animal and human pathogens. Effectiveness of a novel 10 % w/v fipronil spot-on (Eliminall®/Exproline vet™, marketed by Pfizer Animal Health and registered and manufactured by Krka, d.d., Novo mesto) was confirmed against these parasites on experimentally infested cats. Two parallel, unicentre and masked controlled studies were conducted with European mixed breed and mixed sex cats. Cats were allocated randomly to one of two treatment groups based on either pre-treatment flea counts (study 1) or pre-treatment tick counts (study 2). In each of the study, eight animals served as control, while another eight animals were treated once topically with the unit label dose of 50 mg fipronil per cat (10.6-23.8 mg/kg). At each reinfestation, animals were infested with approximately 100 fleas or 60 ticks to achieve adequate infestation rates. Parasites were removed and counted on days 2, 9, 16, 23, 30 and 37, 48 h after the treatment or experimental infestation. Excellent effectiveness was demonstrated on day 2 (100 and 94 % efficacy against fleas and ticks, respectively) and lasted for up to 5 weeks (efficacy ≥96 %) against fleas and up to 4 weeks against ticks (efficacy ≥94 %). The product was well tolerated and no adverse reactions were observed.

The effects of a pour-on formulation of fluazuron 2.5 % and flumethrin 1 % on populations of Rhipicephalus decoloratus and Rhipicephalus microplus both on and off bovine (Bonsmara breed) hosts.

The present study demonstrated the efficacy of a pour-on formulation of fluazuron 2.5 % and flumethrin 1 % (Drastic Deadline eXtreme®) against Rhipicephalus decoloratus and Rhipicephalus microplus on cattle on pasture previously grazed by experimentally infested animals. Six tick-free cattle were placed on the pasture and treated 7 days later (Day 0) with the pour-on. They were retreated on Days 63, 126 and 189 and monthly tick counts were done. Mean numbers of adult R. decoloratus and/or R. microplus decreased from 53 and 14 on Days 56 and 112 respectively to 2 or less on all other occasions including Day 254. Compared to the numbers of R. decoloratus and/or R. microplus larvae collected from vegetation in the previous year, larval numbers declined by 40.7 % on Day 28, and thereafter reduction remained between 84 % and 100 %. Pairs of tracer calves placed on the pasture for 7 days each month were then held in pens and adult ticks that detached collected. Reduction in the numbers of R. decoloratus collected from tracer animals was 75 % on Day 56 and remained above 93 % except for Day 224 when it temporarily decreased to 78.5 %. Reduction in the numbers of R. microplus was 97.5 % on Day 28 and remained above 98 % until the conclusion of the study on Day 254. Treatment with the pour-on formulation of fluazuron and flumethrin resulted in a marked decrease in the numbers of R. decoloratus and/or R. microplus on treated cattle followed by a reduction in the numbers of larvae questing on the vegetation and ticks picked up by tracer calves. No other potential host species for R. decoloratus and/or R. microplus were present in the camps.

Meta-analysis of heritability estimates and genome-wide association for tick-borne haemoparasites in African cattle.

The control of tick-borne haemoparasites in cattle largely relies on the use of acaricide drugs against the tick vectors, with some vaccination also being used against selected pathogens. These interventions can be difficult in Africa, where accessibility and cost of vaccines can be issues, and the increasing resistance of tick vectors to the widely used acaricides is a complication to disease control. A potential complementary control strategy could be the exploitation of any natural host genetic resistance to the pathogens. However, there are currently very few estimates of the extent of host resistance to tick-borne haemoparasites, and a significant contributing factor to this knowledge gap is likely to be the difficulty of collecting appropriate samples and data in the smallholder systems that predominate livestock production in low- and middle-income countries, particularly at scale. In this study, we have estimated the heritability for the presence/absence of several important haemoparasite species (including Anaplasma marginale, Babesia bigemina, Babesia bovis, and Ehrlichia ruminantium), as well as for relevant traits such as body weight and body condition score (BCS), in 1,694 cattle from four African countries (Burkina Faso, Ghana, Nigeria, and Tanzania). Heritability estimates within countries were mostly not significant, ranging from 0.05 to 0.84 across traits and countries, with standard errors between 0.07 and 0.91. However, the weighted mean of heritability estimates was moderate and significant for body weight and BCS (0.40 and 0.49, respectively), with significant heritabilities also observed for the presence of A. marginale (0.16) and E. ruminantium (0.19). In a meta-analysis of genome-wide association studies (GWAS) for these traits, two peaks were identified as reaching the suggestive significance threshold (p < 1.91 × 10-7 and p < 1.89 × 10-7, respectively): one on chromosome 24 for BCS and one on chromosome 8 for the E. ruminantium infection status. These findings indicate that there is likely to be a genetic basis that contributes to pathogen presence/absence for tick-borne haemoparasite species, which could potentially be exploited to improve cattle resistance in Africa to the economically important diseases caused by these pathogens.

Tick-borne pathogens and body condition of cattle in smallholder rural livestock production systems in East and West Africa.

BACKGROUND: The majority of the African population lives in rural areas where they heavily depend on crop and livestock production for their livelihoods. Given their socio-economic importance, we initiated a standardized multi-country (Benin, Burkina Faso, Ghana, Nigeria, Ethiopia Tanzania and Uganda) surveillance study to assess the current status of important tick-borne haemoparasites (TBHPs) of cattle. METHODS: We assessed pathogen prevalences (Anaplasma marginale, Anaplasma centrale, Babesia bigemina, Babesia bovis, Ehrlichia ruminantium, and Theileria parva) in the blood of 6447 animals spread over fourteen districts (two districts per country). In addition, we screened for intrinsic (sex, weight, body condition) and extrinsic (husbandry, tick exposure) risk factors as predictors of infections with TBHPs. RESULTS: There was a large macro-geographic variation observed in A. marginale, B. bigemina, B. bovis and E. ruminantium prevalences. Most correlated with the co-occurrence of their specific sets of vector-competent ticks. Highest numbers of infected cattle were found in Ghana and Benin, and lowest in Burkina Faso. While T. parva was seldomly found (Uganda only: 3.0%), A. marginale was found in each country with a prevalence of at least 40%. Babesia bovis infected individuals had lower body condition scores. Age (as estimated via body weight) was higher in A. marginale infected cattle, but was negatively correlated with B. bigemina and E. ruminantium prevalences. Ehrlichia ruminantium infection was more often found in males, and A. marginale more often in transhumance farming. High levels of co-infection, especially the combination A. marginale × B. bigemina, were observed in all countries, except for Uganda and Burkina Faso. Babesia bigemina was more or less often observed than expected by chance, when cattle were also co-infected with E. ruminantium or A. marginale, respectively. CONCLUSIONS: Tick-borne pathogens of cattle are ubiquitous in African's smallholder cattle production systems. Our standardized study will help a wide range of stakeholders to provide recommendations for TBHP surveillance and prevention in cattle, especially for B. bovis which heavily impacts production and continues its spread over the African continent via the invasive Rhipicephalus microplus tick.

Tick communities of cattle in smallholder rural livestock production systems in sub-Saharan Africa.

BACKGROUND: The majority of the African population lives in rural areas and depends on agriculture for their livelihoods. To increase the productivity and sustainability of their farms, they need access to affordable yield-enhancing inputs of which parasite control is of paramount importance. We therefore determined the status of current tick species with the highest economic impact on cattle by sampling representative numbers of animals in each of seven sub-Saharan countries. METHODS: Data included tick species' half-body counts from approximately 120 cattle at each of two districts per country, collected four times in approximately 1 year (to include seasonality). Study sites were chosen in each country to include high cattle density and tick burden. RESULTS: East Africa (Ethiopia, Uganda and Tanzania) showed overall a higher diversity and prevalence in tick infestations compared to West African countries (Benin, Burkina Faso, Ghana and Nigeria). In East Africa, Amblyomma variegatum (vector of Ehrlichia ruminantium), Rhipicephalus microplus (Babesia bovis, B. bigemina, Anaplasma marginale), R. evertsi evertsi (A. marginale) and R. appendiculatus (Theileria parva) were the most prevalent tick species of economic importance. While the latter species was absent in West Africa, here both A. variegatum and R. microplus occurred in high numbers. Rhipicephalus microplus had spread to Uganda, infesting half of the cattle sampled. Rhipicephalus appendiculatus is known for its invasive behaviour and displacement of other blue tick species, as observed in other East and West African countries. Individual cattle with higher body weights, as well as males, were more likely to be infested. For six tick species, we found reduced infestation levels when hosts were treated with anti-parasiticides. CONCLUSIONS: These baseline data allow the determination of possible changes in presence and prevalence of ticks in each of the countries targeted, which is of importance in the light of human-caused climate and habitat alterations or anthropogenic activities. As many of the ticks in this study are vectors of important pathogens, but also, as cattle may act as end hosts for ticks of importance to human health, our study will help a wide range of stakeholders to provide recommendations for tick infestation surveillance and prevention.

World Association for the Advancement of Veterinary Parasitology (WAAVP) second edition: Guideline for evaluating the efficacy of parasiticides against ectoparasites of ruminants.

This second edition guideline was prepared to assist in the planning, conduct and interpretation of studies to assess the efficacy of parasiticides against ectoparasites of ruminants. It provides updated information on the selection of animals, dosage determination, dosage confirmation and field studies, record keeping and result interpretation. This guideline is intended to assist investigators on how to conduct specific studies, to provide specific information for registration authorities involved in the decision-making process, to assist in the approval and registration of new ectoparasiticides, and to facilitate the worldwide adoption of standard procedures.

World Association for the Advancement of Veterinary Parasitology (WAAVP): Second edition of guidelines for evaluating the efficacy of anthelmintics for dogs and cats.

These revised guidelines have been developed to assist in the design, execution, and interpretation of studies to assess the efficacy of anthelmintic drugs against internal parasites in dogs and cats. The design and execution of studies are outlined and discussed. Considerations for specific targeted parasites are included. Information is provided on the principles of selection of animals, procedures for randomization, housing, feeding, necropsy procedures, and record keeping for dose determination dose confirmation studies and field studies. Complementary to the WAAVP general anthelmintic guidelines, these species-specific guidelines should assist investigators in the evaluation of anthelmintic drugs in dogs and cats by using comparable and standardized procedures in studies with appropriate numbers of animals.

Laboratory breeding of two Phortica species (Diptera: Drosophilidae), vectors of the zoonotic eyeworm Thelazia callipaeda.

BACKGROUND: Some species of drosophilid flies belonging to the genus Phortica feed on ocular secretions of mammals, acting as biological vectors of the zoonotic eyeworm Thelazia callipaeda. This study describes an effective breeding protocol of Phortica variegata and Phortica oldenbergi in insectary conditions. METHODS: Alive gravid flies of P. oldenbergi, P. variegata and Phortica semivirgo were field collected in wooded areas of Lazio region (Italy) and allowed to oviposit singularly to obtain isofamilies. Flies were maintained in ovipots (200 ml) with a plaster-covered bottom to maintain high humidity level inside. Adult feeding was guaranteed by fresh apples and a liquid dietary supplement containing sodium chloride and mucin proteins, while larval development was obtained by Drosophila-like agar feeding medium. The breeding performances of two media were compared: a standard one based on cornmeal flour and an enriched medium based on chestnut flour. All conditions were kept in a climatic chamber with a photoperiod of 14:10 h light:dark, 26 ± 2 °C and 80 ± 10% RH. RESULTS: From a total of 130 field-collected Phortica spp., three generations (i.e. F1 = 783, F2 = 109, F3 = 6) were obtained. Phortica oldenbergi was the species with highest breeding performance, being the only species reaching F3. Chestnut-based feeding medium allowed higher adult production and survival probability in both P. oldenbergi and P. variegata. Adult production/female was promising in both species (P. oldenbergi: 13.5 F1/f; P. variegata: 4.5 F1/f). CONCLUSIONS: This standardized breeding protocol, based on controlled climatic parameters and fly densities, together with the introduction of an enriched chestnut-based feeding medium, allowed to investigate aspects of life history traits of Phortica spp. involved in the transmission of T. callipaeda. Obtaining F3 generation of these species for the first time paved the road for the establishment of stable colonies, an essential requirement for future studies on these vectors in controlled conditions.

A community approach for pathogens and their arthropod vectors (ticks and fleas) in cats of sub-Saharan Africa.

BACKGROUND: Arthropod-borne pathogens and their vectors are present throughout Africa. They have been well studied in livestock of sub-Saharan Africa, but poorly studied in companion animals. Given their socioeconomic importance, the African Small Companion Animal Network (AFSCAN), as part of the WSAVA Foundation, initiated a standardized multi-country surveillance study. METHODS: In six countries (Ghana, Kenya, Nigeria, Tanzania, Uganda, and Namibia) in both rural and urban settings, 160 infested cats were sampled to assess their ectoparasite community (ticks and fleas), as well as the micro-parasite prevalence within those ectoparasites (60 and 118 pools of ticks and fleas, respectively) and blood (276 cats, including 116 non-infested). RESULTS: Almost two thirds of all infested cats originated from Tanzania and Kenya. Despite the large macro-geographical variation, no consistent difference was found in ectoparasite diversity and numbers between East and West Africa. Far more flea-infested than tick-infested cats were found. The most dominant ectoparasite was Ctenocephalides felis. Among the ticks, the exophilic Haemaphysalis spp. were the commonest, including species that are not typically linked with companion animals (Haemaphysalis spinulosa and Haemaphysalis elliptica). The most prevalent pathogens found in the blood and fleas were Bartonella henselae and Mycoplasma haemofelis. In the ticks, the dog-associated Hepatozoon canis was most commonly found. A high degree of co-parasitism was found in all countries and habitats. CONCLUSIONS: Our continent-wide standardized field study highlights the cat's potential to serve as a reservoir of pathogens that can be transmitted to humans or livestock, especially when cats are expected to become more commonly kept in African villages and towns.