Metabolic disruptions in Biomphalaria glabrata induced by Heterorhabditis bacteriophora HP88: Implications for entomopathogenic nematodes in biological control.

Research on the use of entomopathogenic nematodes (EPNs) as a potential tool for the biological control of invertebrates has been growing in recent years, including studies involving snails with One Health importance. In this study, the effect of exposure time (24 or 48 h) of Heterorhabditis bacteriophora HP88 on the activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST), as well as the concentration of total proteins, uric acid, and urea in the hemolymph of Biomphalaria glabrata, were investigated. The concentrations of these metabolic markers were measured weekly until the end of the third week after exposure. Along with a significant reduction in total protein levels, a significant increase (p < 0.01) in uric acid and urea contents in the hemolymph of B. glabrata exposed to H. bacteriophora was observed. The accumulation of urea in these mollusks could lead to deleterious effects due to its high toxicity, inducing significant cell damage. Variations in transaminase activities were also observed, with snails exposed to EPNs showing significantly higher values (p < 0.01) than individuals in the control group, both for ALT and AST. These results indicate that experimental exposure to infective juveniles of H. bacteriophora causes significant alterations in the metabolic pattern of B. glabrata, compromising the maintenance of its homeostasis. Finally, exposure for 48 h caused more damage to the planorbid in question compared to snails exposed for 24 h, suggesting that the exposure time may influence the intensity of the host's response.

What is the optimal timing to initiate strategic control of Rhipicephalus microplus in taurine cattle in a tropical region?

Cattle tick control poses a significant challenge for livestock in tropical and subtropical regions. The objective of this study was to determine the most suitable timing to initiate a strategic tick control program and to identify the ideal number of acaricide treatments for adult taurine cattle (Bos taurus taurus) in a tropical region throughout the year. Three groups with 10 bovines each were performed: T01 (strategic treatment in late autumn/winter/late spring, every 28 days), T02 (strategic treatment to act in the "first tick generation" - early spring/summer/early autumn, every 28 days) and T03 (control). Tick counts (females 4-8 mm) were conducted every 14 days. If the tick burden in any group reached 30 or more during these counts, we applied an additional treatment. Over the course of a year, T02 required significantly fewer (p < 0.05) acaricide treatments than T01, with nine treatments for T02 and eleven for T01. Furthermore, during the tick counts, animals in T02 showed a lower tick burden compared to those in T01. Initiating the strategic tick control program in early spring, corresponding to the first tick generation, proved more effective than starting in autumn. This approach not only required fewer acaricide treatments but also resulted in a reduced tick burden. These benefits are particularly valuable in terms of animal welfare and managing acaricide resistance issues.

Effect of a preventive strategic control program, with imidocarb dipropionate, against tick fever agents in dairy calves.

Chemoprophylaxis with dipropionate imidocarb (IMD) is a method adopted to prevent cattle tick fever (TF). Sixty weaned dairy heifers (±60 days old), without previous exposure to Rhipicephalus microplus ticks, were housed in Tifton paddocks and were subsequently exposed to R. microplus ticks and monitored up to 315 days old. Thirty animals were kept as controls (T01) and 30 received five preventive strategic treatments with IMD at 21-day intervals (T02). The heifers were monitored weekly by means of packed cell volume (PCV) and blood smears to evaluate the presence of TF agents. Salvage treatments (ST) with diminazene and enrofloxacin were administered when animals showed PCV ≤ 24%. The A. marginale prevalence was 39.3% and 37.7%, B. bovis 6.0%, and 7.3%, and B. bigemina 16.3% and 13.7% for T01 and T02, respectively. Regarding PCV values, group T01 showed lower PCV than group T02, between 119 and 161 days of life, but when animals were 196, 210, 217, and between 252 to 301 days old, an inversion occurred. The IMD treatment protocol was effective in group T02 from day 91 to 175 while treatment was being administered, but from day 182 to 315 after the IMD treatment protocol was completed, the number of salvage treatments against TF agents performed in T02 group increased significantly. The sequential application of IMD treatments with intervals less than 21 days is not recommended.

An update on amitraz efficacy against Rhipicephalus microplus after 15 years of disuse.

Amitraz is an acaricide that is widely used in veterinary medicine to control the cattle tick Rhipicephalus microplus. However, controversy exists in the literature regarding the resistance of R. microplus to this product. The present work provides an update on the acaricidal efficacy of amitraz (Triatox®, 12.5 % amitraz) after 15 years without its use on a property. Two in vivo (bovines treated with amitraz and submitted to tick counts, n = 20 animals) and one in vitro (adult immersion test, n = 40 ticks) assays were performed to determine product efficacy. The efficacy of the commercial formulation tested in the first in vivo trial ranged from 14.1 to 47.0%, and in the second from 3.6 to 35.1%, for the 28 days of the experiments. Efficacy for the in vitro trial was 47.38%. The dose recommended by the manufacturer of the product did not cause mortality to most of the ticks of this strain, and efficacy/resistance was not reverted or modified after 15 years (estimated 60 tick generations).

Acaricidal potential of essential oils on Rhipicephalus linnaei: Alternatives and prospects.

The acaricidal potential of various essential oils (EOs) has been evaluated based on their benefits in tick control. This study aimed to investigate the tick-killing activity of Pogostemon cablin "patchouli," Cymbopogon martinii "palmarosa," and Cymbopogon flexuosus "lemongrass" EOs on Rhipicephalus linnaei. Engorged females were collected from domiciled and non-domiciled dogs from Jataí city, Goiás state, to obtain larvae and nymphs in a controlled environment. Two commercial EOs brands were used in this study in different EOs concentrations (2.5, 5, 10, and 20 mg/mL), and was tested by immersion of larvae and nymphs. In the in vitro evaluation of EOs toxicity against R. linnaei larvae, 100 % mortality was achieved with 10 mg/mL of P. cablin oil, whereas mortality rates greater than 98 % were observed with 20 mg/mL of C. martinii and C. flexuosus. In nymphs, high sensitivity was observed, with 100 % mortality achieved using 5 mg/mL of P. cablin and 20 mg/mL of C. martinii and C. flexuosus. The EO of P. cablin "patchouli" demonstrated in vitro toxicity at a lower concentration than the other oils in the two development stages of R. linnaei, and was considered the most efficient and with verified acaricidal activity. Oils of C. martinii "palmarosa" and C. flexuosus "lemongrass" achieved mortality greater than 95 % in larvae and nymphs only at the highest concentration. Therefore, the acaricidal effects of the tested EOs are promising, especially of patchouli oil, which promoted high mortality at a low concentration (LC90 of 2.21 mg/mL).

Annual number of generations and biology of non-parasitic phase of Rhipicephalus microplus in irrigated and non-irrigated pasture in a tropical region.

This study aimed to verify the number of R. microplus annual generations in irrigated and non-irrigated pastures compared to the constant ideal environment. It also sought to evaluate the biology of the non-parasitic phase of this tick for each generation in these different areas of pasture; assess the larvae population dynamics in the pasture of each tick generation, and evaluate the R. microplus population dynamics parasitizing cattle in non-irrigated pasture. In the field experiment, two sub-areas were subjected to artificial irrigation (IRRI-A and IRRI-B) with artesian water, while the other two remained non-irrigated (NIRRI-A and NIRRIG-B). When more than 75 % of the total surviving engorged females from all 90 repetitions of each area (irrigated or non-irrigated) produced mature larvae within one tick generation, two cattle were infested with approximately 10,000 R. microplus larvae from the tick colony used in this study. On the 22nd day post-infestation, a new tick generation was started by releasing these females in different areas (IRRI-B and NIRRIG-B). This procedure was repeated successively, and each year was analyzed independently. In both the non-irrigated and irrigated areas, there were five generations of R. microplus per year. It can be observed that there the number of annual generations of ticks in this region has increased when compared to 30 years ago. Under the constant ideal temperature and humidity conditions (B.O.D. chamber), R. microplus completed an average of 6.59 generations. In the environment, the longest generation was the first (July to October), while the 2nd, 3rd and 4th (December to March) were the most similar to B.O.D. conditions. Although the number of generations was the same in the different areas, the population density of R. microplus larvae was higher in the irrigated area, probably because the irrigation provided milder temperatures, higher relative humidity and lower saturation deficit values during about eight hours per day. Between the 3rd and 5th generation of ticks, there was an overlap of larvae in the pastures, belonging to different generations, and at each peak of infestation observed in cattle between these generations, there were up to 30 % of larvae from the previous generation, and consequently up to 70 % of larvae from the new generation.

A new approach to characterization of the resistance of populations of Rhipicephalus microplus (Acari: Ixodidae) to organophosphate and pyrethroid in the state of Minas Gerais, Brazil.

The monitoring of resistance of cattle tick populations in Brazil to the chemical bases in use is largely limited to investigation of the phenotypic profile. There are few studies investigating the role played by the genotypic profile in acaricide resistance in the country. Therefore, the aim of the present study was to carry out molecular characterization and trace out the genetic profile of populations of Rhipicephalus microplus with respect to resistance to the organophosphate and pyrethroid chemical groups. For that purpose, larvae were genotyped belonging to 587 populations for pyrethroids and 306 for organophosphates, using the polymerase chain reaction technique. It was found that 75.49% and 97.44% of the larvae studied showed resistance to the organophosphates and pyrethroids, respectively. Among the populations resistant to pyrethroids, 91.9% were heterozygotes, showing that most of the resistant populations have only one allele responsible for resistance. Therefore, it is possible to conclude that the genotyped populations have high resistance to organophosphates, and even more so to pyrethroids. This information is fundamental for understanding the mechanisms of resistance of R. microplus to acaricides, to enable improvement of control techniques.

Population dynamics of Rhipicephalus microplus in dairy cattle: influence of the animal categories and correlation with milk production.

Is well known the taurine and zebuine susceptibility to Rhipicephalus microplus. Few are the reports regarding tick population dynamics between the same herd/breed, and because of this, two experiments were performed. In the 1st, the cattle tick population dynamics in dairy nursing calves (reared collective and individually), weaning calves (4-16 months), heifers (17-29 months), cows in lactation and dry cows (≥ 30 months) from the same herd, tick burden and milk production correlation were performed, for two years. R. microplus females (4.5-8.0 mm) counts and the milk production were performed every 28 and 14 days, respectively. In the 2nd experiment, bovines belonging to different categories/age (newborn without previous contact with tick; 12-13 months with tick contact since birth; and 23-24 months with tick contact since birth) were experimentally infested with 30,000 R. microplus larvae, to quantify the number of fully engorged females detached from these animals. In the 1st experiment, when the mean counts of tick were ≥ 30 all animals of the group were treated. Nursing calves showed 3-4 peaks of ticks, animals reared individually showed smaller (p ≤ 0.05) tick burden than those reared collectively. Weaning calves (4-8 months) showed 5 tick peaks/year and higher mean tick burden was found than other categories. On the other hand, animals with 17-29 months of age showed smallest (p ≤ 0.05) tick burden, with 3 tick peaks/year. When the animal become lactating the tick burden increase, and 5 peaks/year occurred, and decrease again in dry cows (p ≤ 0.05) showing 4-5 tick peaks/year. Weaning calves and lactating cows received more acaricide treatments (p ≤ 0.05), 18 and 15, respectively. Nursing calves reared individually, and heifers (21-29 months) were the categories that received two acaricide treatments. The more milk the cow produce, more ticks it has (p ≤ 0.05). In the 2nd experiment, more (p ≤ 0.05) fully engorged females were recovered from younger animals than older ones. So, different tick control strategies need to be adopted in different dairy cattle categories, and the tick burden should be considered, once the effect may be more inherent to the animal rather than the strategy adopted.

Selective versus strategic control against Rhipicephalus microplus in cattle: A comparative analysis of efficacy, animal health, productivity, cost, and resistance management.

This study compared selective control versus strategic control against Rhipicephalus (Boophilus) microplus, under conditions of a real dairy farm. Strategic control requires that all animals be treated with acaricide at regular pre-determined intervals. Selective control only requires treatment of infested animals and only when they are at or above a pre-determined threshold. Tick counts on animals and in pasture were performed and the susceptibility of tick populations to the different treatment methods was evaluated at the beginning and end of the study using the Larval Packet Test, Larval Immersion Test, and Adult Immersion Test. Over the four years of the study strategic control was more advantageous than the selective control as the group experienced lower tick burden on animals and in pasture, absence of skin lesions and myiases, lower operating costs, treatments concentrated in fewer months of the year and lower resistance pressure.

Susceptibility of Amblyomma sculptum, Vector of Rickettsia rickettsii, Ticks from a National Park and an Experimental Farm to Different Synthetic Acaricides.

Amblyomma sculptum is a relevant tick species from a One Health perspective, playing an important role as a vector of Rickettsia rickettsii, the main agent of spotted fever rickettsiosis in Brazil. In this study, we evaluated the susceptibility of two A. sculptum populations from Goiás state (midwestern Brazil) to different acaricides. The first tick population (GYN strain) originated from an experimental farm, where the ticks are annually exposed to acaricides. The second (PNE strain) was collected in a national park (Emas National Park), where the ticks had not been exposed to acaricides. Immersion tests were conducted with 21-day-old laboratory-reared larvae and nymphs originating from adult ticks collected in the areas mentioned above. The chosen acaricides were two synthetic pyrethroids (cypermethrin and deltamethrin), one organophosphate (chlorfenvinphos), one formamidine (amitraz), and two combinations of pyrethroids and organophosphates (cypermethrin, chlorpyrifos and citronellal; cypermethrin, fenthion and chlorpyrifos). Mortality data were used to determine the lethal concentration (LC) values at which 50%, 90%, and 99% of the ticks died (LC50, LC90, and LC99, respectively), and resistance ratios (RR) were calculated based on the LC values. The RR revealed differences between the acaricide-exposed (GYN) and unexposed (PNE) tick strains. The PNE strain larvae and nymphs were susceptible to all the tested acaricides. The GYN strain larvae were tolerant to cypermethrin, whereas the nymphs were tolerant to deltamethrin, chlorfenvinphos, and the combination of cypermethrin, chlorpyrifos, and citronellal (2 < RR ≤ 10). The GYN strain nymphs were resistant to amitraz (RR > 10). This is the first report of A. sculptum nymphs with resistance to amitraz and tolerance to deltamethrin, chlorfenvinphos, and the combination of cypermethrin, chlorpyrifos, and citronellal.

Toxicity of solvents and surfactants to Amblyomma cajennense (Fabricius, 1787) (Acari: Ixodidae) and Dermacentor nitens (Neumann, 1897) (Acari: Ixodidae) larvae.

The objective of this work was to verify the sensitivity of Amblyomma cajennense and Dermacentor nitens larvae to the solvents ethanol, methanol, acetone, xylol and dimethyl sulfoxide (DMSO) and the surfactant Tween 80. The first four solvents were tested at analytical purity while the DMSO and surfactant Tween 80 were tested at a concentration of 1%. The substances tested at analytical purity that caused high mortality were also tested at concentrations of 50%, 25% and 1%. The larval packet test was used, with 10 repetitions for each treatment. A control group was also formed with the same number of repetitions, in which the larvae were only exposed to distilled water. In the first experiment, only xylol was highly toxic at the concentrations tested, causing mortality above 90% for larvae of both species. In the second experiment, xylol at 1% and at 25% showed low toxicity to the A. cajennense and D. nitens larvae, respectively, since the percentage mortality was statistically similar to that of the control group (p>0.05).

Chemical composition and acaricidal activity of essential oil from Lippia sidoides on larvae of Dermacentor nitens (Acari: Ixodidae) and larvae and engorged females of Rhipicephalus microplus (Acari: Ixodidae).

The aim of this work was to identify the compounds and to investigate the acaricidal activity of the essential oil from the leaves of Lippia sidoides on Rhipicephalus microplus and Dermacentor nitens. The oil was obtained by hydrodistillation and analyzed by gas chromatography (GC/FID) and gas chromatography/mass spectrometry. In total, 15 compounds comprising 99.97 % of the total peak area were identified. The main constituent of the essential oil was thymol (67.60 %). The acaricidal activity was assessed by the modified larval packet test, with oil concentrations of 2.5, 5.0, 10.0, 15.0, and 20.0 µl/ml, and by the female immersion test with concentrations of 10.0, 20.0, 40.0, 60.0, and 80.0 µl/ml. The mortality of the R. microplus and D. nitens larvae was greater than 95 % starting at concentrations of 10.0 and 20.0 µl/ml, respectively. In the test with the engorged females, the L. sidoides essential oil starting at a concentration of 40.0 µl/ml caused a significant reduction (p < 0.05) in the values of the egg mass weight and egg production index. The viability of the eggs was affected in all the treated groups, with significantly lower hatching rates (p < 0.05) in relation to the control group. The control percentages at concentrations of 10.0, 20.0, and 30.0 µl/ml were 54, 57, and 72 %, and reached 100 % at the highest two concentrations (60.0 and 80.0 µl/ml). Therefore, it can be concluded that the essential oil from the leaves of L. sidoides has acaricidal activity on R. microplus and D. nitens.

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.

Rhipicephalus microplus seasonal dynamic in a Cerrado biome, Brazil: An update data considering the global warming.

The present study evaluated over two years the seasonal dynamics of Rhipicephalus microplus in a Cerrado biome of midwestern Brazil (Goiânia, Goiás) and correlated the current climatic conditions (environmental temperature, insolation, rainfall, relative humitidy and saturation deficit of the atmosphere) of this site with 30 years ago. In addition, the data collected in the present research were compared with climatic conditions data and R. microplus population dynamics conducted by our group in other regions (Formiga, Minas Gerais and Jaboticabal, São Paulo) within the same biome. For the parasitic phase dynamics, R. microplus females (4.5-8.0 mm) counts were performed on tick natural infested cows kept in Panicum maximum paddocks. To verify the larvae dynamics on pasture the flannel dragg technique on natural infested pasture was performed, and the height of the grass was measured. Five peaks of engorged R. microplus females on animals and larvae infestations on pastures were observed. Being three and two peaks during the rainy and dry season of the year, respectively. Insolation (r = 0.8758; P = 0.00009; R²â€¯= 0.7670), rainfall (r = -0.8572; P = 0.0002; R²â€¯= 0.7348), maximum environmental temperature in summer (r = 0.9999; P < 0.0087; R²â€¯= 0.9988) and the saturation deficit of the atmosphere in autumn (r = -0.9789; P = 0.0211; R²â€¯= 0.9582) influenced the seasonal dynamics of R. microplus larvae on pastures. While the forage height did not influence the number of larvae found on pasture (r = 0.1545; P = 0.7682; R²â€¯= 0.0239). The comparison between the climatic data of the last 30 years with the current climatological data (2012 to 2019) in the state of Goiás, and the comparison with R. microplus population dynamics from other two locations in the same biome indicated that the increase of the environmental temperature due to the global warming possibly is a determining factor which increase the number of one or up two tick annual peaks. In addition, this climatic variable was responsible for increase the population density of ticks on pastures and animals observed in July/winter of the present study, in comparison to other regions with the same biome, as discussed in this work.

Evaluation of different strategic control protocols for Rhipicephalus microplus on cattle according to tick burden.

The development of a strategic protocol is fundamental to the control of Rhipicephalus microplus. The efficacy of different strategic tick control protocols (Day 1 to Day 119) using commercial acaricides according to R. microplus burden on bovines was evaluated. Female ticks (4.5-8 mm in length) were initially counted on cattle to form four groups of bovines with low and high tick burdens (treated and control), with 10 animals each. The animals were then (Day 0) kept in different paddocks, and the bovines from the two treated groups received their first treatment. The main difference in the protocols was the use of a spray formulation on animals with high tick burden. Additionally, the criterion for animal re-treatment (treatment interval) during the proposed strategic protocol was evaluated according to tick length by visual inspection - if 30% of the bovines of a group presented ticks < 4 mm between their legs and dewlap, the treatment was applied, which could change the originally proposed intervals. The number of days for which animals from the treated groups remained without acaricide administration (palliative control, ≥30 tick females < 4 mm) after the end of the strategic tick control protocol was recorded. Tick counts and visual inspection were done weekly until days 203 and 119, respectively. The results showed that the adopted protocol was successful based on calculated efficacy. Mean efficacies from Day 7 to Day 119 were 86.1% and 96.1% for treated groups with low and high tick burdens, respectively, and mean tick count did not differ between them (P > 0.05) but did differ from their respectively controls (P = <0.0001). The use of spray formulation at the beginning of the protocol for animals with a high tick burden helped to avoid harmful tick infestations. Visual analysis of tick length anticipated the proposed treatment intervals. Treated groups remained without palliative treatment for 70 days, while control groups needed treatment every 28 days. It was concluded that strategic control protocols must be developed according to tick burden, and tick length can be used to indicate when treatment needs to be applied.

Heterorhabditis amazonensis (Rhabditidae: Heterorhabditidae), strain RSC-5, for biological control of the cattle tick Rhipicephalus (Boophilus) microplus (Acari: Ixodidae).

The aim of this study was to evaluate the influence of different doses of Heterorhabditis amazonensis RSC-5 on the biological parameters of engorged females of Rhipicephalus (Boophilus) microplus. The female ticks, individually identified, were divided into six groups of 20 each and exposed to the following nematode concentrations: 0, 75, 150, 300, 600, and 1,200/female. The following parameters were observed: initial weight, final weight, alteration weight, egg mass weight, pre-oviposition period, oviposition period, survival period, incubation period, hatching percentage, reproductive efficiency index (REI), nutritional efficiency index (NI), and percentage of control. There were no significant differences observed for the initial weight, pre-oviposition period, incubation period, and NI (p > 0.05) between the various treated groups and the control group. However, there were significant differences (p < 0.01) for the oviposition and survival periods between all the treated groups and the control group. For the final weight, alteration weight, egg mass weight, and REI, only the treatment with 1,200 nematode/female did not differ significantly from the respective figures for the control group (p > 0.05). The highest control rate was 67.8%, obtained in the treatment with a concentration of 300 entomopathogenic nematodes/female. The results demonstrate that this nematode has a deleterious effect on the majority of the biological parameters analyzed.

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.

Potential of entomopathogenic nematodes of the genus Heterorhabditis for the control of Stomoxys calcitrans (Diptera: Muscidae).

The present study verified the pathogenic potential of entomopathogenic nematodes (EPNs) of the genus Heterorhabditis (Heterorhabditis bacteriophora, isolate HP88 and Heterorhabditis baujardi isolate LPP7) to immature stages of Stomoxys calcitrans in the laboratory. All EPN concentrations of the H. bacteriophora HP88 strain caused mean larval mortality greater than 90% after four days. Higher concentrations of the H. baujardi LPP7 isolate (≥50 EPNs/larva) eliminated more than 70% of larvae after six days with the concentration 200 EPNs/larva reaching mortality levels of 93.3%. The larval mortality at all concentrations of EPNs (25, 50, 100, 150, and 200 EPNs/larva) for both strains was significant (p<0.05) when compared to the respective control groups. Concentrations of H. bacteriophora HP88 yielded an LC50 of 0.36 EPN/larva and LC90 of 29.1; while H. baujardi LPP7 yielded an LC50 of 39.85 and LC90 of 239.18. H. bacteriophora HP88 provided greater inhibition of the emergence of adults when compared to the response obtained with H. baujardi LPP7. EPNs did not cause considerable mortality when applied directly to pupae. The set of observed results suggests that the EPNs of the genus Heterorhabditis, isolates HP88 and LPP7, are a promising alternative in the control of the stable fly.

Entomopathogenic nematodes in insect cadaver formulations for the control of Rhipicephalus microplus (Acari: Ixodidae).

This study evaluated the efficacy of four entomopathogenic nematode (EPN) strains in insect cadaver formulations against Rhipicephalus microplus and compared the efficacy of the most virulent EPNs applied in cadavers of Galleria mellonella and Tenebrio molitor. In the first experiment, infected G. mellonela larvae were used as the source of EPNs. Engorged females of R. microplus were placed in pots filled with soil and different numbers of G. mellonella larvae infected with one of four species of nematodes. All treatments with EPNs of the genus Heterorhabditis caused significant reduction (p<0.05) in the egg mass weight and hatching percentage of larvae. The EPNs of the genus Steinernema, except for the group exposed to Steinernema carpocapsae ALL, whose source nematodes included six larvae of G. mellonella, caused a significant reduction (p<0.05) in the egg mass weight produced per female. Steinernema feltiae SN applied with two, four, and six cadavers and S. carpocapsae ALL with two cadavers caused a reduction in hatching percentage of larvae of R. microplus (p<0.05). The percentage of control was above 95% in all groups treated with Heterorhabditis bacteriophora HP88 and Heterorhabditis indica LPP1 and in the treatment with four larvae infected with S. feltiae SN. The second experiment followed the same methodology, using G. mellonella and T. molitor larvae infected by the two most virulent EPNs. H. bacteriophora HP88 and H. indica LPP1 in different formulations caused reduction in the egg mass weight and hatching percentage of larvae. The percentage of control were 82.4 and 84.9% for H. bacteriophora HP88 and H. indica LPP1, respectively, formulated in T. molitor, and reaching 99.9% in groups formulated with G. mellonella. The EPNs tested in insect cadaver formulation showed pathogenicity to engorged females of R. microplus and EPNs of the genus Heterorhabditis formulated in G. mellonella larvae were more effective.