Toltrazuril + fenbendazole for cattle: Pharmacokinetics and efficacy against Eimeria spp. and gastrointestinal nematodes.

The present work evaluated the pharmacokinetics and efficacy of the association of 15cmg/kg toltrazuril +5cmg/kg fenbendazole against Eimeria spp. and gastrointestinal nematodes (GINs) in calves of different regions of Brazil (Center-West, Southeast, and South). A total of seven experiments were carried out, five of which determined formulation efficacy against Eimeria spp., considering the following aspects: therapeutic, preventive, metaphylactic, and residual efficacy. Therapeutic efficacy experiments for GINs were carried out by parasitological necropsy. The toltrazuril + fenbendazole association demonstrated ≥95% efficacy against Eimeria spp. for 21 days post-treatment (DPT). When used preventively and metaphylatically, the same association demonstrated ≥97% efficacy against E. zuernii, E. ellipsoidalis, E. cylindrica, E. bovis, E. wyomingensis and E. auburnensis. Toltrazuril + fenbendazole administered seven days before challenge was 100% effective against all these Eimeria species. Results of therapeutic, preventive, metaphylactic and residual efficacies can be related to the pharmacokinetic results, especially considering toltrazuril sulfone, which was detected in animal plasma for a longer period than the parent compound. Toltrazuril + fenbendazole achieved 100% anthelminthic efficacy against the GINs Haemonchus placei (L4), Cooperia pectinata and Oesophagostomum radiatum; 99.94% against adult H. placei; and 99.98% against C. puntacta. The association of toltrazuril + fenbendazole, associated with other measures, is an important and suitable tool for the control and treatment of Eimeria spp. and GINs in young cattle.

Tick infestation level interferes with spray formulation (organophosphate + pyrethroid) efficacy against Rhipicephalus microplus.

Due to the scarcity of information regarding the control of Rhipicephalus microplus on bovines with different tick burdens, tick counts and the efficacy of a commercial spray formulation (aspersion bath with organophosphate + pyrethroid) were evaluated for animals with a low and high tick burdens for 35 days in a stall test. Thirty-two crossbreed Bos taurus indicus x Bos t. taurus experimentally infested with R. microplus larvae were divided into four groups: T01 high infestation control, T02 high infestation treated, T03 low infestation control, and T04 low infestation treated. The bovines were kept individually in pens and R. microplus females were collected daily. All data were statistically analyzed with a significance level of 5% (P ≤ 0.05). For linear regressions, variables with P≤0.05 and the highest coefficients of determination (R2 ≥ 0.70) were considered the best descriptors. The reliability level was 95%. Tick counts differed (P<0.0001) between T01 and T02 (high infestation control vs. treated), except at day 2 (P >0.05) post-treatment, and efficacy ranged from 85.2 to 50.6%. The number of collected engorged females was lower (P<0.0001) for T04 than for T03 (low infestation treated vs. control) animals from day 3 to 29, and efficacy ranged from 95.2 to 69.8%. In addition, tick burden and efficacy were negatively correlated for both the group of animals with a high tick burden (r = -0.5256; p = 0.0012; R² = 0.2762) and the group of animals with a low tick burden (r = -0.9817; p < 0.0001; R² = 0.9638). In conclusion, a high tick burden on bovines decreases the efficacy of the tested spray acaricide.

Strategic control of cattle co-parasitized by tick, fly and gastrointestinal nematodes: Is it better to use ecto + endoparasiticide or just endectocide formulations?

Ticks, flies, and gastrointestinal helminths (GINs) significantly affect cattle productivity; thus, ectoparasiticide, endoparasiticide, and endectocide drugs have commonly been used for their control. The study aimed to compare the technical (parasites counts), productive, and financial effects of a treatment protocol comprising ecto- + endoparasiticides formulations (T01: fluazuron 2.5 mg/kg + fipronil 1.25 mg/kg and fenbendazole 5 mg/kg; n = 15) to a treatment with one formulation of endectocide (T02: ivermectin 450 µg/kg + abamectin 250 µg/kg; n = 15) over 308 days under field conditions in crossbred cattle co-parasitized by Rhipicephalus microplus, Haematobia irritans, and GINs. Bovine weight gain and return on investment (ROI) were also evaluated. Bovines from T01 received four treatments against the cattle tick and two against two GINs. For T02, four treatments were performed. Animals from T01 gained 15.4 kg more than T02 and provided a comparative ROI of 15.8. In cattle co-parasitized with R. microplus, H. irritans, and GINs, the treatment protocol used in this study with ecto- + endoparasiticidal action formulations showed better technical results regarding parasite counts and productive and financial data than the strategic treatment protocol using only an endectocide formulation.

Investigation of fecal egg counts versus worm burden and helminth fauna in cattle treated or not with macrocyclic lactones in a tropical region.

Gastrointestinal nematodes negatively impact the health and productivity of livestock. Macrocyclic lactones (MLs) are the most common class of chemicals used in helminth control programs, however, their intense use is accelerating the development of parasite resistance. For cattle, little is known regarding the relationship between fecal egg counts (FECs) and the helminth population inside the animal's gastrointestinal tract and so this study evaluated the correlation between FEC and worm burden in cattle treated or not with MLs (ivermectin 200 µg/kg, ivermectin 630 µg/kg, abamectin 200 µg/kg, doramectin 700 µg/kg and moxidectin 200 µg/kg). Animals were necropsied to determine if there were any modifications to the proportions of worm species due to chemical treatment. FECs of ML-treated and untreated animals showed significant (p ≤ 0.05 and R2 ≥0.70) positive linear correlations and substantial or perfect strength-of-agreement (LCCC ≥ 0.61) with total worms present, all of which were Haemonchus placei and Cooperia punctata. However, this correlation and strength-of-agreement did not occur when the efficacy of MLs was ≥80 % against these helminths. Among the active MLs tested, moxidectin had the weakest relationship between FEC and worm burden, except for H. placei. Analysis of the helminthological fauna found an increase of H. placei (35.6 % = 95 % CI 35.4-35.6) and a consequent decrease of C. punctata (52.5 % = 95 % CI 52.3-52.6) in untreated animals, when compared to studies carried out over 20-40 years ago, but this proportion was reversed for ML-treated animals (C. punctata 64.5 % = 95 % CI 64.4-64.8; H. placei 30.8 % = 95 % CI 30.6-30.8). It is possible that MLs mitigate this effect in the field and that C. punctata remained prevalent in cattle in different regions of Brazil.

Does bathing affect tick and flea burdens and ectoparasiticide effectiveness of a spot-on formulation (fipronil + (S)-methoprene) for dogs?

This work evaluated tick and flea burdens and the efficacy of a single topical application of 10% fipronil + 9% (S)-methoprene spot-on against experimental infestations of Rhipicephalus sanguineus sensu lato and Ctenocephalides felis felis on dogs that were submitted to regular baths for 56-days post-treatment. Four treatments (n = 24) were evaluated: no chemical treatment and no bathing (T01); no chemical treatment + bathing (T02); chemical treatment and no bathing (T03) and chemical treatment + bathing (T04). Dogs were infested with adult ticks and fleas seven days pre-treatment (-7). The ectoparasites were then counted and removed on day -5, followed by a new infestation on day -2 and treatment on day 0. Ticks and fleas were then counted without removal on day 1 and counted with removal on day 2. The dogs were then repeatedly infested with both ectoparasites on days 5, 12, 19, 26, 33, 40, 47 and 54 post-treatment. Baths and ectoparasite counts with removal were performed every 48 h before and after experimental infestation, respectively. Animals of T04 had lower (P ≤ 0.05) tick burden than animals from T01 and T02 from day 2 to day 28. For fleas, the counts varied among days of the study. Dogs from T03 presented lower (P ≤ 0.05) flea burden than dogs from T01 from day 1 until day 56. The same was true for T04 when compared to T01 from day 1 to day 42. Animals of T02 greater (P ≤ 0.05) flea burden than animals of T04 in all dates post treatment. In addition, dogs from T02 presented higher (P ≤ 0.05) flea burden than T01 from day 7 to day 28 and from day 49 to day 56. In sum, the efficacy of the spot-on formulation used declined over the time against both ectoparasites. The efficacy for ticks ranged from 0.0 to 96.5% and 0.0 to 98.52% for T03 and T04, respectively. While for fleas, the efficacy of the formulation ranged from 67.41 to 100% for T03 and 40.18 to 100% for T04. So, bathing dogs that were not treated increased C. f. felis burden but not R. sanguineus s. l. burden. The residual efficacy of the product had a shorter duration against these ectoparasites for dogs that received subsequent bathing.

Biological parameters for Rhipicephalus microplus in the field and laboratory and estimation of its annual number of generations in a tropical region.

The present study aimed to evaluate biological parameters of Rhipicephalus microplus in the non-parasitic phase in both field and laboratory conditions. It also aimed to assess correlations between duration (in days) of these parameters and climatic variables (humidity, rainfall, and soil temperature) and to estimate the annual number of generations of R. microplus in a tropical region. The non-parasitic phase of R. microplus in field and laboratory conditions was evaluated throughout the course of two years. A pasture was infested with engorged female of R. microplus, and biological parameters, including female pre-oviposition, female oviposition, egg mass incubation, larval pre-hatching phase, larval maturation, and larval longevity, were evaluated concomitantly with the collection of data on climatic conditions. The same parameters were also evaluated in a climatized chamber in the laboratory. The total duration of the non-parasitic phase in the field was longer in the dry season (1st and 4th life-cycle repetitions) than in the rainy season (2nd, 3rd, and 5th repetitions). Tick biological parameters for the non-parasitic phase in the laboratory were similar to those obtained in the field during the rainy season. The evaluated biological parameters were influenced mainly by environmental and ground-level temperatures, which modified egg mass incubation, larval pre-hatching, and larval longevity periods and, consequently, the total duration of the non-parasitic phase of the tick. The annual number of generations for the tick was estimated at five per year, which is alarming because it represents an increase, and so new studies into strategic control are needed.

In vivo effect of diflubenzuron, administered via mineral salt supplementation, against Haematobia irritans and Rhipicephalus microplus parasitizing cattle.

This study involved two field trials with the aim of evaluating the efficacy of diflubenzuron, via mineral supplementation, against Haematobia irritans parasitizing cattle. Concomitantly with the main trial, a stall test was conducted to ascertain the effects of a different formulation with the same active ingredient against Rhipicephalus microplus, along with the action of diflubenzuron on the reproductive parameters of R. microplusfemales that had naturally detached from cattle. Against H. irritans, it was observed that the efficacy indexes fordiflubenzuron were low (≤ 31.3% or 44.6%) or null (0.0%) throughout the study. The anti- R. microplus efficacy of diflubenzuron, at weekly intervals, ranged from 0.0 to 13.7% over the entire experimental period. Null efficacy (0.0%) was registered for diflubenzuron in relation to the reproductive parameters of R. microplusfemales that had naturally detached from cattle. The different diflubenzuron formulations, administered via mineral salt supplementation, did not show satisfactory efficacy indexes against H. irritans and R. microplus parasitizing cattle, within the experimental design of the present study. In addition, this agent did not present any deleterious effects on the reproductive parameters of R. microplus females.

In vivo effect of diflubenzuron, administered via mineral salt supplementation, against Haematobia irritans and Rhipicephalus microplus parasitizing cattle / Efeito in vivo do diflubenzuron, administrado via sal mineral, contra Haematobia irritans e Rhipicephalus microplus parasitando bovinos

Abstract This study involved two field trials with the aim of evaluating the efficacy of diflubenzuron, via mineral supplementation, against Haematobia irritans parasitizing cattle. Concomitantly with the main trial, a stall test was conducted to ascertain the effects of a different formulation with the same active ingredient against Rhipicephalus microplus, along with the action of diflubenzuron on the reproductive parameters of R. microplusfemales that had naturally detached from cattle. Against H. irritans, it was observed that the efficacy indexes fordiflubenzuron were low (≤ 31.3% or 44.6%) or null (0.0%) throughout the study. The anti- R. microplus efficacy of diflubenzuron, at weekly intervals, ranged from 0.0 to 13.7% over the entire experimental period. Null efficacy (0.0%) was registered for diflubenzuron in relation to the reproductive parameters of R. microplusfemales that had naturally detached from cattle. The different diflubenzuron formulations, administered via mineral salt supplementation, did not show satisfactory efficacy indexes against H. irritans and R. microplus parasitizing cattle, within the experimental design of the present study. In addition, this agent did not present any deleterious effects on the reproductive parameters of R. microplus females.

Resumo O objetivo deste estudo foi avaliar a eficácia do diflubenzuron, administrado via suplementação mineral, contra Haematobia irritans parasitando bovinos, em dois testes à campo. Concomitantemente, foi realizado testes em estábulo para determinar os efeitos de uma formulação diferente, com o mesmo princípio ativo, contra Rhipicephalus microplus , bem como a ação do diflubenzuron nos parâmetros reprodutivos de fêmeas de R. microplus recolhidas após desprendimento natural do hospedeiro bovino. Contra H. irritans, foi observado que foram baixos (≤ 31,3% ou 44,6%) ou nulos (0,0%) os índices de eficácia do diflubenzuron. A eficácia anti-R. microplus do diflubenzuron, observada em intervalos semanais, variaram de 0,0% a 13,7% durante todo o período experimental. Com relação aos parâmetros reprodutivos das fêmeas de R. microplus recolhidas, foi observada eficácia nula (0,0%) para o diflubenzuron. Conclui-se que as diferentes formulações administradas via sal mineral no atual estudo, contra H. irritans e R. microplus parasitando bovinos, não apresentaram eficácia satisfatória. Este agente também não mostrou efeito deletério sobre os parâmetros reprodutivos de fêmeas de R. microplus.

Prevalence of bovine fascioliasis, areas at risk and ensuing losses in the state of Goiás, Brazil / Prevalência de fasciolose bovina, áreas de risco e perdas subsequentes no estado de Goiás, Brasil

Abstract The present study had to determine the prevalence and spatial distribution of areas at risk of bovine fascioliasis in the state of Goiás, central-western Brazil between 2007 and 2014; to evaluate the associations of some epidemiological variables with occurrences of Fasciola hepatica in animals; and to estimate the economic losses that this parasite on the cattle industry. It could be concluded that of 23,255,979 animals slaughtered, the average prevalence of F. hepatica in cattle in Goiás during the period evaluated was 0.0026% (95% CI: 0.0024-0.0028). In the State of Goias, in about then years (since when this parasite was found for the first time by other researchers in 2007), F. hepatica was diagnosed in 168 new municipalities. Using the regression analysis, the effective bovine herd size was a significant risk factor (OR= 1.21; 95% CI 1.1022-1.4510; p ≤ 0.05) for cattle to be infected with fascioliasis in the state of Goiás. The cattle-rearing industry lost approximately R$ 15,072.75 (US$ 4,785) due to condemn of livers with Fasciola in the state of Goiás. New studies need to be conducted in these regions, with the aim to identify the likelihood of presence of intermediate hosts, which might serve as a source of F. hepatica infection for definitive hosts.

Resumo O presente estudo objetivou determinar a prevalência e a distribuição espacial das áreas em risco para fasciolose bovina no Estado de Goiás, região Centro-Oeste do Brasil, entre 2007 a 2014; avaliando as associações de variáveis epidemiológicas com ocorrências da Fasciola hepatica em animais; e estimar as perdas econômicas deste parasito em bovinos. Pode-se concluir que de 23.255.979 bovinos abatidos, a prevalência média de F. hepatica durante o período avaliado foi de 0,0026% (IC 95%: 0,0024-0,0028). No Estado de Goiás, em aproximadamente 10 anos (desde quando este parasito foi encontrado pela primeira vez por outros pesquisadores em 2007), F. hepatica foi diagnosticada em 168 novos municípios. Utilizando-se a análise de regressão logística, o tamanho do rebanho apresentou-se como um fator de risco significativo (OR= 1,21; IC 95% 1,1022-1,4510; p≤0,05) para os bovinos se infectarem por F. hepatica no referido Estado. O prejuízo, devido à condenação de fígados com Fasciola nos bovinos no Estado de Goiás, foi de R$ 15.072,75 (US 4.785). Novos estudos precisam ser conduzidos nessas regiões, com o objetivo de identificar a presença de hospedeiros intermediários infectados, que podem servir como fonte de infecção por F. hepatica para hospedeiros definitivos.

Prevalence of bovine fascioliasis, areas at risk and ensuing losses in the state of Goiás, Brazil.

The present study had to determine the prevalence and spatial distribution of areas at risk of bovine fascioliasis in the state of Goiás, central-western Brazil between 2007 and 2014; to evaluate the associations of some epidemiological variables with occurrences of Fasciola hepatica in animals; and to estimate the economic losses that this parasite on the cattle industry. It could be concluded that of 23,255,979 animals slaughtered, the average prevalence of F. hepatica in cattle in Goiás during the period evaluated was 0.0026% (95% CI: 0.0024-0.0028). In the State of Goias, in about then years (since when this parasite was found for the first time by other researchers in 2007), F. hepatica was diagnosed in 168 new municipalities. Using the regression analysis, the effective bovine herd size was a significant risk factor (OR= 1.21; 95% CI 1.1022-1.4510; p ≤ 0.05) for cattle to be infected with fascioliasis in the state of Goiás. The cattle-rearing industry lost approximately R$ 15,072.75 (US$ 4,785) due to condemn of livers with Fasciola in the state of Goiás. New studies need to be conducted in these regions, with the aim to identify the likelihood of presence of intermediate hosts, which might serve as a source of F. hepatica infection for definitive hosts.

Avermectin toxicity in bovines less than thirty days old.

The present study was designed to evaluate possible adverse effects of different dosages of avermectins (abamectin and a combination of ivermectin + abamectin) administered subcutaneously in calves less than one month of age. Clinical as well as biochemical parameters of blood and cerebrospinal fluid (CSF) were evaluated for animals subjected to various treatments. Thirty-five calves were divided into seven groups of five animals each: T01 (control), T02 (abamectin 200 µg/kg), T03 (abamectin 400 µg/kg), T04 (abamectin 600 µg/kg), T05 (ivermectin 450 µg/kg + abamectin 250 µg/kg), T06 (ivermectin 900 µg/kg + abamectin 500 µg/kg) and T07 (ivermectin 1350 µg/kg + abamectin 750 µg/kg). Cerebrospinal fluid and blood samples were collected on day zero (treatment day) and on days 1, 7 and 14 following treatment. The biochemical parameters measured in blood samples were total protein and the AST, GGT and alkaline phosphatase enzymes. In CSF samples, color, aspects, pH, density, cellularity, total proteins, glucose and the CK, ALT and LDH enzymes were evaluated. No neurological toxicity or biochemical changes in the CSF or blood that could be correlated to administration of tested products were observed in this study. Though it is well known that abamectin may lead to intoxication in bovines less than four months of age, based on data from the present study, no adverse clinical effects to the CSF or blood were observed in calves <30 days old that had received up to 600 µg/kg abamectin formulations and up to 1350 µg/kg ivermectin +750 µg/kg abamectin associations. It is possible that the level and activity of the multiple drug resistance gene (MDR1 - ABCB1) present in the calves may have influenced the results obtained in this study. Future studies should be carried out.

Susceptibility of Rhipicephalus (Boophilus) microplus to fluazuron (2.5mg/kg) and a combination of novaluron (2.0mg/kg)+eprinomectin (0.36mg/kg) in field studies in Brazil.

The present study aimed to determine the susceptibility of 32 R. (B.) microplus populations from Southeast, Midwest and South regions of Brazil, to fluazuron (2.5mg/kg), administered topically (pour-on). Additionally, five populations (Southeast and Midwest regions) of the southern cattle tick were evaluated using in vivo field studies, regarding their susceptibility to a new combination of novaluron (2.0mg/kg)+eprinomectin (0.36mg/kg), administered subcutaneously, compared with two positive controls (fluazuron 2.5mg/kg and eprinomectin 0.5mg/kg), both administered topically (pour-on). Selected bovines were allocated to treatment groups on day 0, and block formation was based on arithmetic means of female ticks (4.5-8.0mm long) counted on three consecutive days (-3, -2 and -1). To evaluate therapeutic and residual efficacies of these formulations, tick counts (females ranging from 4.5 to 8.0mm long) were performed on days 3, 7 and 14 post-treatment, continuing on a weekly basis until the end of each experiment. Results obtained throughout this study, utilizing field efficacy trials, allowed us to conclude that four R. (B.) microplus populations (including two in the Southeast and two in the Midwest regions) could be diagnosed as resistant, or with low susceptibility, to fluazuron (2.5mg/kg). Such fact was detected in farms where owners applied products containing this active component on cattle for at least five years, with treatment intervals of 30-55days during the rainy season. Nonetheless, in vitro studies should be performed in order to reinforce in vivo results obtained on the present study. Regarding efficacy indexes obtained by the association of eprinomectin and the novel molecule novaluron against R. (B.) microplus, none of the trials managed to obtain efficacies superior to 48%. Such results, allied to data obtained by different researchers and previously published in literature, reinforce the perception that maybe these formulations containing novaluron, in the administered dosages and treatment routes, may not be effective tools for controlling R. (B.) microplus. However, future studies must be conducted in order to support such hypothesis. Additionally, all five R. (B.) microplus populations were diagnosed as resistant, or with low susceptibility, to eprinomectin (0.5mg/kg) as well. Even though fluazuron, administered topically (pour on), is still an excellent active principle to be used against R. (B.) microplus, resistance management strategies should be quickly implemented in order to keep selection pressure in Brazil at a minimum level for this compound.

Population dynamics and evaluation of the partial selective treatment of crossbreed steers naturally infested with Rhipicephalus (Boophilus) microplus in a herd from the state of Minas Gerais in Brazil.

The present study aimed to evaluate the population dynamics of Rhipicephalus (Boophilus) microplus over a period of 13 months on a rural property located in the state of Minas Gerais in southeastern Brazil. Animals were treated for ticks indoors by whole body spraying when R. (B.) microplus had an average count equal or more than 30 ticks. The study also evaluated the possibility of a partial selective treatment for bovines to control R. (B.) microplus in which only a percentage of the population would be treated (specifically those bovines with tick counts of ≥20. Moreover, we examined the percentage of the population of R. (B.) microplus present on experimental bovines that did not come into contact with the chemical compounds used in the partial selective treatment. We concluded that in this particular region of Brazil, the crossbreed steers support up to five R. (B.) microplus generations per year and that the number of generations was primarily affected by the pluviometric precipitation. We sprayed the bovines with chemicals seven times during the course of the study. The results of the partial selective treatment method revealed that during the rainy and the dry periods, 42.1% to 60.0% and 61.9% to 79.2% of the animals, respectively, fulfilled the criteria to receive a chemical treatment to reduce the number of cattle ticks. In consideration of the need to slow the development of tick resistance with the chemical compounds used in the spraying treatment, the results showed that the percentage of animals that did not require treatment is not relevant. This was evidenced by the result that bovines that presented tick counts of ≥20 during the dry and rainy periods represented 91.5% and 90.6% of the total recorded R. (B.) microplus populations, respectively. Only 8.7% of the tick population remained free from exposure to acaricides during the 13 months of the study, which is an important point when considering the adoption of the partial selective treatment method. Future studies with larger herds must be conducted on these topics; however, our results suggest that the partial selective treatment method most likely will not slow the development of resistance in this R. (B.) microplus population against the chemical compound used in this study.

Ten years later: Evaluation of the effectiveness of 12.5% amitraz against a field population of Rhipicephalus (Boophilus) microplus using field studies, artificial infestation (Stall tests) and adult immersion tests.

Using field trials, artificial infestations (Stall tests) and in vitro adult immersion tests, the present study evaluated the acaricidal efficacy of 12.5% amitraz administered via whole body spraying against a Rhipicephalus (Boophilus) microplus population that did not have any contact with chemical products belonging to this acaricide family for 10 years (approximately 40 generations). Two natural infestation trials, two artificial infestation trials (Stall tests) and two adult immersion tests were performed in two different stages in 2005 and 2015. Between 2002 and 2015, the bovine herd of this property was formed by approximately 450 animals from the Simmental breed that were divided into nine paddocks formed by Cynodon dactylon (L.) Pers. For the natural infestation experiments in 2005 and 2015, we selected nearly 70 animals naturally infested with ticks from the same herd that belonged to the "São Paulo" farm located in São José do Rio Pardo, São Paulo, Brazil. Field studies were performed in the same paddock (9). To evaluate anti-R. (B.) microplus activity in the artificially infested cattle (Stall tests) and adult immersion tests, two experiments of each methodology were performed at CPPAR (the Center of Research in Animal Health located on the FCAV/UNESP campus in Jaboticabal, São Paulo, Brazil) in 2005 and 2015. R. (B.) microplus used in the artificial infestation, and adult immersion test experiments were obtained from paddocks 1-9 in 2005 and 2015 from the commercial farm where the field studies were performed. Based on the obtained results, it was possible to conclude that amitraz use in rotation with pyrethroids every 28 days for three consecutive years (2002-2004) previous to the beginning of the first trial (2005) was sufficient to generate a R. (B.) microplus strain resistant to amitraz. Moreover, using field trials, artificial infestations (Stall tests) and adult immersion tests, we verified that 40 generations of the tick species with no contact to the aforementioned compound (amitraz) were not sufficient to revert or modify the efficacy/resistance of amitraz for this analyzed R. (B.) microplus strain. The reversion of amitraz efficacy values in R. (B.) microplus may only occur when resistance of the field strain is incipient. Alternatively, the differences in the results may be due to differences in the Rhipicephalus spp. species between current study locations. Therefore, future studies must be performed to prove this hypothesis.

Effects of different spray formulations on the reproductive parameters of engorged Rhipicephalus (Boophilus) microplus females detached from experimentally infested cattle.

This present study aimed to evaluate the deleterious effects of some commercially available spray formulations (15% Cypermethrin+25% Chlorpyriphos+1% Citronellal and 8% Cypermethrin+60% Ethion) on the reproductive parameters of engorged Rhipicephalus (Boophilus) microplus females that detached from experimentally infested cattle. The following reproductive parameters of engorged female ticks were analyzed: female weight, egg mass weight, percentage of hatchability, percentage of reduction in oviposition, percentage of reduction in hatchability, reproductive efficiency and percent control/efficacy of formulations for reproductive parameters. Our findings showed that although the strain R. (B.) microplus used in both experiments was thought to be sensitive to the test compounds because of the acaricidal efficacy observed throughout these trials, it was not possible to observe overall deleterious effects on the reproductive parameters of this tick species with both spray formulations. However, the 8% Cypermethrin+60% Ethion showed short-term significant effects on the weight of female ticks between the 14th and 16th days post-treatment and the weight of female and the egg mass weight between the 20th and 22nd days post-treatment. New studies should be conducted to show if these results regarding the reproductive parameters of fully engorged R. (B.) microplus females, combined with the acaricidal efficacy can be sufficient to reduce the number of chemical treatments administered to cattle.

Susceptibility of helminth species from horses against different chemical compounds in Brazil.

By means of parasitological necropsies, the present study aimed to evaluate, in six experiments, the degree of susceptibility or resistance of different helminth species which naturally infect horses to ivermectin 0.2 mg/kg, abamectin 0.2 mg/kg, moxidectin 0.4 mg/kg, trichlorfon 35 mg/kg, ivermectin 0.2 mg/kg+praziquantel 2.5 mg/kg, abamectin 0.2 mg/kg+praziquantel 2.5 mg/kg and ivermectin 0.2 mg/kg+6.6 mg/kg pyrantel. At experimental day zero, the horses were allocated to treatment groups based on average counts of strongylid eggs per gram of feces (EPG) obtained on days -3, -2 and -1. Oxyuris sp. infections were confirmed as positive or negative. All the animals in the six experiments were naturally infected by this helminth species. Each group (control or treated) consisted of six animals. All the assessed Habronema muscae populations analyzed were susceptible to ivermectin, abamectin and moxidectin. Of the six Trichostrongylus axei populations, four were susceptible to ivermectin, abamectin, moxidectin, trichlorfon and ivermectin+praziquantel, and two were resistant to abamectin+praziquantel and ivermectin+pyrantel. Both Strongyloides westeri populations analyzed were susceptible to ivermectin, abamectin, moxidectin and abamectin+praziquantel. For O. equi, resistance was found in four different populations treated with ivermectin, abamectin, moxidectin, trichlorfon and ivermectin+praziquantel. Only combinations of abamectin+praziquantel and ivermectin+pyrantel were effective against this parasite species. All the large strongyles diagnosed in the present study (Strongyus edentatus, Strongyus vulgaris and Triodontophorus serratus) were susceptible to all the chemicals tested, with the exception of trichlorfon. Of the Cyathostominae populations, one was diagnosed as resistant to ivermectin and another to trichlorfon. The remaining populations from this nematode group were considered to be sensitive to ivermectin, abamectin, moxidectin, ivermectin+praziquantel, abamectin+praziquantel and ivermectin+pyrantel. New studies should be performed in different regions to evaluate the efficacy of trichlorfon in others field populations of helminthes.

Acaricidal effects of fluazuron (2.5 mg/kg) and a combination of fluazuron (1.6 mg/kg) + ivermectin (0.63 mg/kg), administered at different routes, against Rhipicephalus (Boophilus) microplus parasitizing cattle.

The present study aimed to evaluate the acaricidal efficacy of fluazuron (2.5 mg/kg), administered as a pour-on, in comparison to an injectable formulation containing fluazuron (1.6 mg/kg) + ivermectin (0.63 mg/kg), against Rhipicephalus (Boophilus) microplus in naturally and experimentally infested cattle. Two studies were conducted with different tick strains, one with artificial infestations (Stall Test, using leight animals per group) and one with natural infestations (utilizing ten animals per group). In both studies, the animals were randomized, according to average tick counts performed on days -3, -2 and -1, into four groups: T01, negative control (saline solution); T02, pour-on fluazuron (2.5 mg/kg); T03: subcutaneous fluazuron (1.6 mg/kg) + ivermectin (0.63 mg/kg); and T04 subcutaneous ivermectin (0.63 mg/kg). Based on obtained results, and considering the utilized tick strains, it was possible to conclude that the pour-on fluazuron (2.5 mg/kg) formulation demonstrated high acaricidal efficacy, with protection periods ranging from 49 to 77 days against Rhipicephalus (Boophilus) microplus. On the other hand, for the injectable fluazuron (1.6 mg/kg) + ivermectin (0.63 mg/kg) formulation, it was not possible to observe elevated anti-R. (B.) microplus effect on both artificial and experimental infestation studies. Results observed for this combination were similar or inferior to those obtained by subcutaneous ivermectin (0.63 mg/kg). Future studies with this formulation containing fluazuron (1.6 mg/kg) + ivermectin (0.63 mg/kg), regarding pharmacokinetic and/or bioavailability profiles, or even studies analyzing both this active principles separately, are needed, seeking to better understand the effects of such combination against Rhipicephalus (Boophilus) microplus parasitizing cattle.

A comparison of three different methodologies for evaluating Rhipicephalus (Boophilus) microplus susceptibility to topical spray compounds.

This study aimed to compare three different methodologies (Adult Immersion Tests, field trials with naturally infected animals, and a Stall Test using artificially infested cattle) to evaluate the efficacy of two topical formulations that we administered as whole body sprays (15% Cypermethrin+30% Chlorpyriphos+15% Fenthion-Colosso(®) FC 30, Ouro Fino Agronegócios; and 60% Dichlorvos+20% Chlorpyriphos-Ectofós(®), Vallée Saúde Animal Ltd.), against a susceptible strain of Rhipicephalus (Boophilus) microplus. To achieve this objective, two natural infestation trials were conducted, as well as two artificial infestation trials (Stall Tests) and two Adult Immersion Tests (AIT). The AIT results showed that both spray formulations achieved 100% efficacy against R. (B.) microplus fully engorged females. However, when observing results obtained by field trials (natural infestations) and Stall Tests, none of these topically applied compounds reached 100% efficacy or affected the reproductive capacity of the fully engorged female ticks. Additional studies must be conducted to compare these in vivo methodologies with different in vitro techniques, such as the Larval Packet Test. However, based on results obtained here, we can conclude that depending on the spray formulations used, the AIT can overestimate acaricidal efficacy and values of reproductive efficiency of such compounds against R (B.) microplus. Specifically, when dealing with spray formulations in the Stall Tests, the period of residual action can increase because these animals are sheltered from contact with environmental factors that might interfere with the efficacy of the products tested. It may be necessary to take in vivo trial results into consideration (such as field trials with naturally infested animals or Stall Tests) to standardize a specific in vitro assay, such as the Adult Immersion Test.

Effect of a spray formulation on the reproductive parameters of a susceptible population of Rhipicephalus (Boophilus) microplus.

The number of studies emphasizing the possible damage that acaricidal spray formulations can cause on engorged female ticks' reproductive parameters is small. The present study evaluated the deleterious effects of a spray formulation (dichlorvos 60% + chlorpyrifos 20%) on the reproductive parameters of a susceptible population of Rhipicephalus (B.) microplus females, using the Stall Test. The ticks were allocated randomly to treatments according to the mean numbers of females detached from each cow on days -3, -2 and -1 and the cattle pen location. The numbers of engorged female ticks that naturally detached from the cattle were counted daily from day 1 to day 30. For each group, 20 detached engorged female ticks or the available number collected daily were evaluated regarding reproductive parameters. Associations of organophosphates demonstrated elevated acaricidal efficacy, as well as deleterious effects on the reproductive parameters of R. (B.) microplus females. The engorged female weight (days 1 to 7), weight of egg masses (days 5 to 10) and larval hatching percentage (days 5 to 19) were decreased (P ≤ 0.05). It is possible that a formulation can lead to deleterious effects on R. (B.) microplus females when the tick population analyzed shows elevated sensitivity towards a particular formulation. However, further studies need to be conducted.

Endoparasites of horses from the Formiga city, located in center-west region of the state of Minas Gerais, Brazil.

With the aim of studying the endoparasite fauna of horses from the Formiga city, located in center-west region of the state of Minas Gerais, 25 animals that were naturally infected with helminths were evaluated. By means of parasitological necropsies, different endoparasites were found. The subfamily Cyathostominae presented the highest incidence, followed by Trichostrongylus axei, Oxyuris equi, Triodontophorus serratus, Strongyloides westeri, Strongylus edentatus, Habronema muscae, Parascaris equorum, Probstmayria vivipara, Strongylus vulgaris, Gasterophilus nasalis, Anoplocephala magna and Anoplocephala perfoliata. In the present study, if the species Probstmayria vivipara was not considered in the prevalence, the frequency of Cyathostominae was equivalent to 94.85%. The results obtained in this study allowed us to detect and identify different species of helminths in horses, and confirmed the high incidence of nematodes belonging to the subfamily Cyathostominae in the center-west region of Minas Gerais.