Assessment of weight gain and control of parasitism by Rhipicephalus microplus in dairy cattle in the field using organosynthetic parasiticide, phytotherapics and homeopathy / Avaliação do ganho de peso e controle do parasitismo por Rhipicephalus microplus em bovinos leiteiros a campo usando parasiticida organosintético, fitoterápicos e homeopatia

The goal of this research was to compare the effect of the following products on dairy cattle, parasitized by Rhipicephalus microplus: organosynthetics - Clorfenvinfós and Ivermectina (OG); phytotherapics - Eucalyptus oil (EG) and Neem cake (NG); Homeopathy (HG); The control group (CG) received no treatment. Infestation was by R. microplus (MIC) and weight gain (GP) were evaluated. The study included 60 animals (5 groups) from ­ Dutch / Zebu, aged between 25-44 months and initial weight between 211-477kg. EG and NG showed MIC 84.9% and 14.0% greater than CG, respectively (P <0.05; P = 0). HG and OG had MIC less than CG 24.4% and 16.9%, respectively (P<0.05; P <0.05). EG, NG and OG obtained GP lower than CG in 7.9%, 8.9% and 8.06% (P <0.05) respectively. The HG GP was 4.9% higher (P <0.05) than CG. This was the first research to prove parasitism control by R. microplus in a field test, using homeopathy. The methodology for choosing Homeopathy for the control of enzooties, developed and tested for the first time in this study, proved to be adequate and efficient, opening the possibility for establishing a new methodology for strategic control of parasitism by R. microplus.(AU)

O objetivo desta pesquisa foi comparar o efeito dos seguintes produtos sobre bovinos leiteiros, parasitados por Rhipicephalus microplus: organossintéticos - clorfenvinfós e ivermectina (OG); fitoterápicos - óleo de eucalipto (EG) e torta de Neem (NG) e homeopatia (HG). O grupo controle (CG) não recebeu tratamento. Foram avaliados infestação por R. microplus (MIC) e ganho de peso/grupo (GP) em 60 animais (5 grupos), a partir de ­ holandês/zebu, com idade entre 25 e 44 meses e peso inicial entre 211 e 477kg. EG e NG apresentaram MIC 84,9% e 14,0% maior que CG, respectivamente (P<0,05; P=0). HG e OG apresentaram MIC menor que CG 24,4% e 16,9%, respectivamente (P<0,05; P<0,05). EG, NG e OG obtiveram GP menor que CG em 7,9%, 8,9% e 8,06% (P<0,05), respectivamente. O GP do HG foi 4,9% maior (P<0,05) que o CG. Esta foi a primeira pesquisa a comprovar controle do parasitismo por R. microplus em teste a campo, usando homeopatia. A metodologia para escolha de medicamentos homeopáticos para controle de enzootias, desenvolvida e testada pela primeira vez neste estudo, mostrou-se adequada e eficiente, abrindo a possibilidade para estabelecimento de nova metodologia para controle estratégico do parasitismo por R. microplus.(AU)

Metarhizium anisopliae sensu lato (s.l.) oil-in-water emulsions drastically reduced Rhipicephalus microplus larvae outbreak population on artificially infested grass.

Rhipicephalus microplus Canestrini, 1887 (Arachnida: Ixodidae) is a mandatory bloodsucking ectoparasite, and it is considered one of the main sanitary problems in livestock. In the development of new technologies for controlling ticks, Metarhizium anisopliae complex (M. anisopliae s. l.) Sorokin, 1883 (Ascomycota: Clavicipitaceae) have been shown to be an effective and safe alternative. However, when applied in the field, abiotic factors may directly influence their microbial activity, and different oily adjuvants enhance fungal efficacy, stability and viability under environmental conditions. Here, the efficacy of two Metarhizium anisopliae s.l. isolates (CG 148 and CG 347) were compared in mineral oil (MO) or vegetable oil (VO) emulsions to control cattle ticks under semi-natural conditions, as well as their persistence in the soil over time. Engorged female ticks were placed on the soil of grass pots previously treated or not with conidial oil-in-water emulsions. Both M. anisopliae s.l. oil-in-water emulsions tested were able to control R. microplus, achieving up to 100% daily efficacy, and remained viable in the soil for up to 60 days after treatment. These results show the prospective applicability of Metarhizium spp. to control the cattle tick and a real possibility for its control in the environment.

Nomuraea rileyi as biological control agents of Rhipicephalus microplus tick.

Nomuraea rileyi, a fungus pathogenic to insects, has been widely used for biological control of agricultural pests in Brazil. This study investigates the effects of N. rileyi, isolates Nr 138, Nr 151, and Nr 177, to eggs, larvae, and engorged females of Rhipicephalus microplus tick. Specimens were immersed in 1 ml of conidial suspension for 3 min, whereas the control group was immersed in 0.01% Tween 80 water solution. The isolate Nr 138 controlled 67.37% of ticks when the highest conidial concentration was used, 10(8) conidia ml(-1). The isolate Nr 177 significantly reduced the percentage of hatch of larvae from eggs treated with 10(8) conidia ml(-1). Conversely, the isolate Nr 151 was not virulent to eggs, larvae, or adults. Variability in virulence was observed among the N. rileyi isolates investigated in the current study-Nr 138 was more virulent to engorged females, while Nr 177 was more virulent to unfed larvae. Although N. rileyi proved to be virulent to several stages of R. microplus, the results obtained in this study indicate that N. rileyi does not appear to be a remarkable biological control agent for R. microplus.

Susceptibility of different populations of ticks to entomopathogenic fungi.

This study aimed to evaluate the in vitro effect of the entomopathogenic fungi Metarhizium anisopliae sensu lato (s.l.) and Beauveria bassiana sensu lato (s.l.) on two distinct populations of Rhipicephalus microplus, from two different experimental farms. Bioassays were performed with engorged females, eggs and larvae. Fungal infection was evaluated based on biological parameters of treated engorged females, percentage of hatch from treated eggs, and percentage of mortality and mean lethal time (LT90) of treated larvae. When the treatments were compared between the two populations, there were significant differences in the following parameters: pre-oviposition period, hatching period and egg production index. Moreover, the results showed that B. bassiana s.l., isolate Bb 986, was more virulent than M. anisopliae s.l., isolate 959, for engorged females, showing a control percentage of 49%. In the bioassay with eggs, the hatching percentage ranged from 3.1% to 49.5% in one population and from 3.4% to 42.7% in the other, with no significant difference between the two populations. In the bioassay with unfed larvae, the mortality percentage ranged from 91.8% to 98.7% in one population and from 71.0% to 94.0% in the other. The LT(90) varied from 19.52 to 27.51 days in one of the populations and 22.89 to 37.31 days in the other. These results suggest that populations of R. microplus show distinct variation in their susceptibility to B. bassiana s.l. and M. anisopliae s.l.

Haemolymph protein and lipid profile of Rhipicephalus (Boophilus) microplus infected by fungi.

The current study evaluates the protein and lipid profile of haemolymph of Rhipicephalus (Boophilus) microplus engorged females infected by Metarhizium anisopliae, Beauveria bassiana or Fusarium oxysporum. Ticks were immersed or inoculated with conidial suspension. Haemolymph was collected from the dorsal surface of engorged females. The results showed altered total protein amounts; however, no significant difference was observed on electrophoretic profile among haemolymph samples. In addition, altered lipid profile was detected in haemocyte samples from ticks treated with Beauveria and Metarhizium.

Genetic diversity among Brazilian isolates of Beauveria bassiana: comparisons with non-Brazilian isolates and other Beauveria species.

AIMS: The genetic diversity of Beauveria bassiana was investigated by comparing isolates of this species to each other (49 from different geographical regions of Brazil and 4 from USA) and to other Beauveria spp. METHODS AND RESULTS: The isolates were examined by multilocus enzyme electrophoresis (MLEE), amplified fragment length polymorphism (AFLP), and rDNA sequencing. MLEE and AFLP revealed considerable genetic variability among B. bassiana isolates. Several isolates from South and Southeast Brazil had high similarity coefficients, providing evidence of at least one population with clonal structure. There were clear genomic differences between most Brazilian and USA B. bassiana isolates. A Mantel test using data generated by AFLP provided evidence that greater geographical distances were associated with higher genetic distances. AFLP and rDNA sequencing demonstrated notable genotypic variation between B. bassiana and other Beauveria spp. CONCLUSION: Geographical distance between populations apparently is an important factor influencing genotypic variability among B. bassiana populations in Brazil. SIGNIFICANCE AND IMPACT OF THE STUDY: This study characterized many B. bassiana isolates. The results indicate that certain Brazilian isolates are considerably different from others and possibly should be regarded as separate species from B. bassiana sensu latu. The information on genetic variation among the Brazilian isolates, therefore, will be important to comprehending the population structure of B. bassiana in Brazil.

Ação do fungo Beauveria bassiana associado a gel polimerizado de celulose no controle do carrapato Anocentor nitens em teste de campo / Action of the fungus Beauveria bassiana associated with cellulose polymerized gel in the control of the tick Anocentor nitens in a field test

Foram selecionados 20 equídeos naturalmente infestados por Anocentor nitens, dos quais seus pavilhões auriculares foram submetidos a quatro tratamentos distintos: gel associado a Beauveria bassiana, apenas gel, apenas B. bassiana e um grupo-controle. Procederam-se à contagem e ao registro do número total de fêmeas adultas (>3mm) em cada pavilhão auricular, nos dias 0, 1, 4, 7, 11, 14, 18, 21, 25 e 28, para cálculo do percentual de controle. Foram coletadas e mantidas em laboratório 20 fêmeas pertencentes a cada tratamento, para cálculo dos períodos de pré-postura, postura, incubação e eclosão, e peso médio das posturas. No grupo tratado com o composto gel associado a B. bassiana, observou-se percentual de controle maior que 50 por cento, entre o 4º e o 25º dia, assim como decréscimo progressivo do peso médio da postura das fêmeas coletadas no período entre o 7º e 14º dia. Para os outros tratamentos, o percentual de controle manteve-se abaixo de 20 por cento, e o peso da postura não apresentou diferença significativa. Os percentuais de controle, assim como as reduções no peso médio da postura, sugerem que a utilização do composto gel associado a B. bassiana potencializou a virulência do entomopatógeno.

Twenty horses naturally infected by Anocentor nitens were selected and their auricular pinna were submitted to four treatments: gel associated with Beauveria bassiana, gel only, B. bassiana only, and a control group. The count and the register of the total number of grown up tick females (>3mm) were performed in each auricular pinna on days 0, 1, 4, 7, 11, 14, 18, 21, 25, and 28 for the calculus of percentage of control. Twenty females from each treatment were collected and maintained in laboratory, to calculate the periods of pre-oviposition, oviposition, incubation, and hatching and mean weight of oviposition. In the group treated with the gel associated to B. bassiana, it was observed a control percentage higher than 50 percent from the 4º to the 25º day, such as a progressive decrease of mean weight of the ovipositions in the period from the 7º to 14º day. For the other treatments, the control percentage was kept below 20 percent and the oviposition weight showed no significant difference among the treatments. The control percentages, such as reductions in mean weight of oviposition, suggest that the use of gel associated with B. bassiana potentialized the virulence of the entomopathogen.

Efeitos de Beauveria bassiana (Bals) Vuill e Metarhizium anisopliae (Metsc) Sorok sobre fêmeas ingurgitadas de Amblyomma cajennense (Fabricius, 1787) em condições de laboratório / Effects of Beauveria bassiana (Bals) Vuill and Metarhizium anisopliae (Metsc) Sorok on engorged females of Amblyomma cajennense (Fabricius, 1787) in laboratory conditions

The in vitro susceptibility of Amblyomma cajennense engorged females to some isolated of the fungus Beauveria bassiana and Metarhizium anisopliae was verified and lethal concentrations (LC 50 and LC 90) were calculated. The females were dived in conidia suspensions for five minutes, and kept in climatically controlled chambers BOD under 27º C and 80 por cento relative humidity. Each bioassay had four treatments in concentrations of 105,106,107 e 108 conidia/ml. A control group was also used. The following characteristics were evaluated: weight and period of oviposition, indexes of reproductive and nutritional efficiency and percentage of microbiological control. A dose dependent negative effect was observed in ticks treated with the suspension. All isolates tested cause a negative effect on in vitro tests of engorged females of A. cajennense, suggesting its potential for microbiological control of tick's species.

Isolation of Beauveria bassiana and Metarhizium anisopliae var. anisopliae from Boophilus microplus tick (Canestrini, 1887), in Rio de Janeiro State, Brazil.

The objective of this work were to isolate and identify strains of entomopathogenic fungi from ingurgitated female Boophilus microplus ticks, collected from the soil in the municipality of Paracambi, Rio de Janeiro State, Brazil. The ingurgitated females were inoculated in the selective medium oat dodine agar (oda), where 49 colonies of Beauveria bassiana (71%) and 20 of Metarhizium anisopliae var. anisopliae (29%) were isolated. These isolated strains characterize for the first time in Brazil the natural occurrence of these species of fungi in this tick, and will be used to conduct bioassays to evaluate the pathogenicity and virulence of these strains for ticks of the genus Boophilus microplus.

Açäo in vitro dos fungos Beauveria bassiana (Bals) Vuill e Metarhizium anisopliae (Metsch) Sorok sobre ninfas e adultos de Amblyomma cajennense (Fabricius, 1787) (Acari: Ixodidae) / In vitro of the fungi Beauveria bassiana (Bals) Vuill and Metarhizium anisopliae (Metsch) Sorok on ninphs and adults of Amblyomma cajennense (Fabricius, 1787) (Acari: Ixodidae)

O trabalho teve por objetivo verificar a mortalidade in vitro de ninfas e adultos de Amblyomma cajennense frente à açäo de isolados dos fungos Beauveria bassiana e Metharhizium anisopliae. Foram avaliados três isolados de M. anisopliae (959, 319 e E9) e dois de B. bassiana (986 e 747). As suspensöes de conídios foram preparadas a partir de fungos produzidos em meio de arroz, e cada bioensaio foi constituído de quatro tratamentos nas concentraçöes 10 elevado a 5, 10 elevado a 6, 10 elevado a 7, 10 elevado a 8 conídios/ml e um grupo-controle. A análise constou da observaçäo do percentual de mortalidade, 15 dias após o tratamento ou após a ecdise de adultos. Foram observadas diferenças significativas entre os tratamentos quanto à mortalidade para todos os isolados e todos os instares. Nos grupos tratados houve aumento considerável na mortalidade à medida que se aumentava a concentraçäo de conídios na suspensäo. Conclui-se que todos os isolados testados causaram mortalidade em testes in vitro sobre esses estádios evolutivos, sugerindo o controle do A. cajennense pela açäo desses fungos

Efeito do isolado 986 do fungo Beauveria bassiana (Bals) Vuill sobre fêmeas ingurgitadas de Anocentor nitens Neumann, 1897 (Acari: Ixodidae) / Effect of isolate 986 of the fungi Beauveria bassiana (Bals) Vuill on engorged females of Anocentor nitens Neumann, 1897 (Acari: Ixodidae)

Fêmeas ingurgitadas de Anocentro nitens foram colhidas de eqüinos infestados para determinar a influência do fungo Beauveria bassiana (isolado 986) na fase näo parasitária do carrapato. Grupos de 20 fêmeas foram utilizados para testar suspensöes que continham água destilada, espalhante adesivo e diferentes concentraçöes de conídios (10 elevado a quinta potência, 10 elevado a sexta potência, 10 elevado a sétima potência e 10 elevado a oitava potência conídios/ml), mais um grupo-controle. As fêmeas foram imersas nas suspensöes por três minutos, acondicionadas em placas de Petri e levadas a câmara climatizada (27ºC e 80 por cento UR). Com exceçäo do período de pré-postura e do percentual de eclosäo das larvas, os resultados revelaram diferenças significativas entre os grupos tratados e o controle. Nos grupos tratados obrservou-se diminuiçäo gradativa, inversamente proporcional às concentraçöes de conídios, no período de postura, no peso da massa de ovos, no período médio de sobrevivência, no período de incubaçäo, no período de eclosäo e nos índices de eficiência reprodutiva e nutricional. Contrariamente, houve aumento gradativo, diretamente proporcional às concentraçöes, no peso da quenógina e na mortalidade de larvas. Entretanto, näo foi observada interferência sobre o percentual de eclosäo. Os resultados obtidos permitem concluir que B. bassiana pode ser utilizada em programas de controle biológico de fêmeas de A. nitens