Biomarkers of gastrointestinal nematodes in beef cattle raised in a tropical area.

Biomarkers are specific molecular, histological, or physiological characteristics of normal or pathogenic biological processes and are promising in the diagnosis of gastrointestinal nematodes (GINs). Although some biomarkers have been validated for infection by Ostertagia sp. in cattle raised in temperate regions, there is a lack of information for tropical regions. The aim of this project was to assess potential biomarkers and validate the most promising. In the first study, 36 bovines (Nelore breed) naturally infected by GINs were distributed into two groups: infected (not treated with anthelmintic) and treated (treated with fenbendazole on days 0, 7, 14, 21, 28, 42, and 56). The variables of interest were live weight, fecal egg count, hemogram, serum biochemical markers, phosphorus, gastrin, and pepsinogen. In the second step, pepsinogen was assessed in cattle of the Nelore breed distributed among three groups: infected (not treated with anthelmintic), MOX (treated with moxidectin), and IVM + BZD (treated with ivermectin + albendazole). In the first study, no difference between groups was found for weight, albumin, hematocrit (corpuscular volume [CV]), erythrocytes, or hemoglobin. Negative correlations were found between pepsinogen and both CV and albumin, and albumin was negatively correlated with the percentage of Haemonchus sp. in the fecal culture. Among the biomarkers, only pepsinogen differentiated treated and infected (beginning with the 28th day of the study). In the second study, a reduction in pepsinogen was found after anthelmintic treatment. Therefore, pepsinogen is a promising biomarker of worms in cattle naturally infected by the genera Haemonchus and Cooperia in tropical areas.

Anthelmintic resistance of gastrointestinal nematodes in cattle in Brazil and Argentina - current status and global perspectives.

This review outlines the current state of anthelmintic resistance (AHR) of gastrointestinal nematodes (GINs) among cattle in Argentina and Brazil, emphasizing the economic repercussions, animal health and welfare. The analysis explores factors associated with AHR and proposes a potential solution: the use of drug combinations. Both countries are grappling with a severe AHR scenario in cattle, having progressed through incipient, established, and advanced phases, leading to extreme cases of animal mortality due to ineffective control strategies. Genera such as Cooperia and Haemonchus have the highest reports of resistance, with Oesophagostomum radiatum also posing significant problems. While oral benzimidazoles and levamisole remain effective in most herds, moxidectin is entering an advanced resistance phase, and avermectins are increasingly deemed ineffective. The review explores the impact ofclimate, mixed grazing, animal movement and other husbandry practices, and the relationship between ectoparasite control and the emergence of resistant helminths. Notably, the discussion includes the strategic use of drug combinations as a valuable approach to address resistant GINs control in livestock, highlighting its significant potential to mitigate the challenges posed by AHR in the cattle industry of these countries.

Two protocols using fluralaner for Rhipicephalus microplus strategic control on taurine cattle in a tropical region.

BACKGROUND: The present study aimed to evaluate the effects of different treatment strategies using a new commercial formulation containing pour-on fluralaner on Rhipicephalus microplus infestation in cattle and in pastures in a tropical climate region where up to five generations of this tick species can occur per year. METHODS: Forty-five naturally infested cattle were divided into three experimental groups: T01, treated with fluralaner (2.5 mg/kg) pour-on every 42 days; T02, the cattle received the first treatment with fluralaner on Day 0 but the next treatment involved a weekly visual evaluation; T03, control, received palliative treatment with a spray formulation when the group mean was ≥ 30 ticks. Counts of female R. microplus were performed weekly until day 343, and larval counts on pasture were performed on Days 0, 30, and 60 and every 30 days until Day 330. RESULTS: Using fluralaner, six applications were performed in Group T01, and four were performed in Group T02. In the control group (T03), it was necessary to perform eight palliative acaricide treatments with the spray formulation. The animals in T01 and T02 showed lower mean tick counts (p ≤ 0.05) than the control group (T03) on 28 and 27 of the 49 evaluated dates, respectively. In the paddock where the animals were kept as controls, the R. microplus larvae counts increased to 1458. In the paddocks where the animals were treated with fluralaner, the number was ≤ 19 per paddock during the study. CONCLUSIONS: The different strategic treatment protocols performed with pour-on fluralaner (2.5 mg/kg) over a year in taurine cattle in a tropical region with a history of up to five annual generations of cattle ticks were effective, maintaining levels of R. microplus infestations in animals and in pastures close to zero in most of the study. Depending on the retreatment criterion adopted, the number of applications per year may be lower, resulting in a reduction in the mean cost of acaricide treatment per year and lower exposure of R. microplus populations to the active ingredient, resulting in lower resistance and selection pressure.

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.

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.

Status of Rhipicephalus microplus resistance to ivermectin, fipronil and fluazuron in Mato Grosso do Sul, Brazil.

Southern cattle tick resistance to pour-on and injectable acaricides has yet to be evaluated on a broader scope, and the paucity of information on the subject may hinder efforts to control this parasite. The objective of this study was to evaluate the resistance profile of ten populations of Rhipicephalus microplus to the acaricides fluazuron, fipronil and ivermectin in cattle herds in Mato Grosso do Sul, Brazil. The larval immersion test (LIT) was used to evaluate susceptibility to ivermectin and fipronil and the adult immersion test (AIT) was performed to evaluate fluazuron. Samples were randomly obtained in ten farms, and in general, we found resistance in five samples to fluazuron and in four samples to ivermectin and fipronil. Six samples showed incipient resistance to ivermectin and fipronil. Five of the ten evaluated samples showed resistance and/or incipient resistance to all the active ingredients, and the other five to two active ingredients. Among the samples classified as resistant, the average resistance ratio for ivermectin was 2.75 and 3.26 for fipronil. These results demonstrate the advanced status of resistance to the most modern chemical groups for the control of R. microplus in the state of Mato Grosso do Sul.

First report of Dermatobia hominis resistant to doramectin in cattle.

The objective of this study was to evaluate the therapeutic efficacy of 1 % injectable doramectin to control Dermatobia hominis in naturally infested beef cattle in a farm with history of clinical parasitism after avermectin use. The study was conducted in a commercial beef cattle farm in Brazil. Twenty crossbred cattle (Nellore x Angus) were selected based on the minimum number of 10 live larvae. Assessment of infestation was performed based on tactile-visual inspection of the entire body of the animals. The animals were allocated to two experimental groups of ten animals each: treated group - dosed with doramectin 200 µg/kg live weight (LW), and control group - dosed with saline solution 1 mL/50 kg LW. Both treatments were administered subcutaneously in a single dose. After treatment, larval counts were performed at 1, 3, 7 and 14 days post-treatment (DPT). For the treatment to be considered efficient, it should reach an efficacy greater than or equal to 90 % by the 10th DPT and the average larval count in the treated animals should be statistically lower than the average larval count in the control group. The geometric means of larval counts at the beginning of the study were 27.2 and 26.3 for the control and treated groups, respectively. After 72 h of treatment, there was a significant reduction (p < 0.01) in the larvae counts of the treated group, extending to the 14th DPT, at which point maximum efficacy for doramectin (86.3 %) was obtained. After treatment, 90 % of the animals remained parasitized with live larvae on the 7th DPT and 70 % on the 14th DPT, while in the control group, all animals were infested on all dates. The administration of injectable doramectin was not effective in controlling D. hominis in naturally infested cattle.

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.

Gastrointestinal nematode control programs in yearling Nellore heifers: Analysis of fecal egg counts, weight gain and reproductive indices.

The objective of this study was to evaluate two different gastrointestinal nematode treatment regimens. Fecal egg counts (FECs), proportion of nematode genera, weight gain, body condition score and reproductive indices (estrous cyclicity, conception and pregnancy rates) were evaluated in yearling heifers after imposing two treatment regimens for gastrointestinal nematodes: T1 = 306 calves treated in May and November with 3.5 % doramectin (700 µg/kg) and August (Aug) with saline solution; and T2 = 307 calves treated in May with 3.5 % doramectin (700 µg/kg), in August with 1% moxidectin (200 µg/kg) and in November with 3.5 % doramectin (700 µg/kg). The animals were weighed, and feces were collected for conducting FECs and coproculture. There was imposing of three fixed-time artificial inseminations (TAIs), and estrous cyclic and pregnancy statuses were determined. Cooperia was the most frequent genus detected in both groups. Heifers of the T2, as compared to those in the T1 group, had fewer FECs in November (P ≤  0.05) and greater weight gain and average daily weight gain (P ≤  0.05) from August to November. There tended to be more heifers of the T2 than T1 group estrous cycling (P =  0.07) at the beginning of the breeding season as well as greater pregnancy rates (P =  0.03) and conception rates (P =  0.03) as a result of the second FTAI. The results indicate there is greater reproduction outcomes as a result of strategic control of gastrointestinal nematodes in yearling Nellore heifers using the T1 as compared with T2 treatment regimen.

Clinical safety of lauric acid for cattle and its in vitro and in vivo efficacy against Rhipicephalus microplus.

The aim of the present study was to test the in vitro acaricidal activity of saturated fatty acids (hexanoic, octanoic, decanoic, lauric, myristic, palmitic, octadecanoic, eicosanoic, docosanoic and tetracosanoic) against Rhipicephalus microplus and select a candidate compound for the subsequent determination of its clinical safety for mice and bovines as well as its in vivo efficacy (ethical clearance number 507/2013). None of the compounds exhibited in vitro larvicidal effectiveness, but acaricidal effectiveness was greater than 95 % in the adult immersion test at 40 mg/ml (hexanoic, octanoic, decanoic, lauric, myristic, palmitic and eicosanoic acids). After a second AIT evaluation of serial concentrations of the fatty acids, lauric and myristic acids were selected for the safety and in vivo efficacy assays. No adverse effect was found in the local lymph node assay in mice treated with lauric or myristic acid. Moreover, no clinical signs of systemic poisoning or dermatological, hematological or biochemical abnormalities were found in cattle after the topical application of 1 % lauric acid. In the dose determination test, the 1% solution of this compound exhibited 86% efficacy in cattle naturally infested by a field population of Rhipicephalus microplus susceptible to all chemical groups, except synthetic pyrethroids. The efficacy of 1 % lauric acid was 53.4 % in the dose confirmation test performed on another herd with a field R. microplus population resistant to all chemical groups of acaricides. In conclusion, fatty acids are potential bioactive compounds for the control of R. microplus. Topically applied lauric acid (C12) exhibits in vivo acaricide activity against adults, nymphs and larvae of R. (B) microplus and is safe for cattle.

Combination of quercetin and ivermectin: In vitro and in vivo effects against Haemonchus contortus.

The aim of the present study was to evaluate the in vitro effect of quercetin combined with ivermectin (IVM) on Haemonchus contortus larvae and adults with different resistance profiles and demonstrate the in vivo anthelmintic action of this combination when used in sheep naturally infected. The effect of combination was evaluated based on the analysis of the mean effective concentration (EC50) obtained for larvae using the larval migration inhibition test and for adults using the motility test on females. The tests with larvae and adults were conducted using isolates with different degrees of susceptibility to IVM (sensitive, intermediate and highly resistant). The in vivo effect was evaluated based on the reduction in the egg count (FEC) and reduction in the count of adult helminths recovered after parasitological necropsy. Using the combination of quercetin with IVM, it was observed that in larvae, quercetin did not significantly reduce the EC50 for IVM in the sensitive and highly resistant isolates, but led to a significant reduction in the EC50 for IVM in the intermediate isolate. In adults, quercetin did not significantly reduce the EC50 for IVM in any of the isolates. No significant effect of the combination was found regarding the reduction in FEC or total count of parasites. The results of the in vitro and in vivo tests performed in the present study on quercetin activity underscore the importance of evaluating resistance-reversing agents among different stages of parasite development as well as among isolates with different resistance profiles. The action of quercetin combined with IVM on the motility of H. contortus larvae and adults was influenced by the degree of resistance and development stage of the parasite. The combination was effective only on intermediate resistant larvae. No action of the combination against adults was found. Moreover, this combination, when administered through the intra-abomasal route, was not effective at reducing the FEC and parasite load of naturally infected sheep.

Discovery of potential ovicidal natural products using metabolomics.

Plant extracts are a potential source of new compounds for nematode control and may be an excellent alternative for the control gastrointestinal nematodes that are resistant to conventional anthelmintics. However, research involving natural products is a complex process. The main challenge is the identification of bioactive compounds. Online analytical techniques with universal detectors, such as high-performance liquid chromatography-mass spectrometry (HPLC-MS), together with metabolomics could enable the fast, accurate evaluation of a massive amount of data, constituting a viable option for the identification of active compounds in plant extracts. This study focused on the evaluation of the ovicidal activity of ethanol extracts from 17 plants collected from the Pantanal wetland in the state of Mato Grosso do Sul, Brazil, against eggs of Haemonchus placei using the egg hatchability test. The ethanol extracts were obtained using accelerated solvent extraction. The data on ovicidal activity, mass spectrometry and metabolomics were evaluated using HPLC-DAD-MS, partial least squares regression analysis (PLS-DA) and a correlation map (univariate correlation analyses) to detect compounds that have a positive correlation with biological activity. Among the ten metabolites with the best correlation coefficients, six were phenylpropanoids, two were triterpene saponins, one was a brevipolide, and one was a flavonoid. Combinations of metabolites with high ovicidal action were also identified, such as phenylpropanoids combined with the triterpene saponins and the flavonoid, flavonoids combined with iridoid and phenylpropanoids, and saponins combined with phenylpropanoid. The positive correlation between classes of compounds in plants belonging to different genera and biological activity (as previously identified in the literature) reinforces the robustness of the statistical data and demonstrates the efficacy of this method for the selection of bioactive compounds without the need for isolation and reevaluation. The proposed method also enables the determination of synergism among the classes, which would be impracticable using traditional methods. The present investigation demonstrates that the metabolomic technique was efficient at detecting secondary metabolites with ovicidal activity against H. placei. Thus, the use of metabolomics can be a tool to accelerate and simplify bioprospecting research with plant extracts in veterinary parasitology.

Retrospective study of epidemiological, clinical and pathological findings of bovine babesiosis in Mato Grosso do Sul, Brazil (1995 -2017).

Babesiosis is a tick-borne hemolytic disease caused by intraerythrocytic protozoan parasites of the genus Babesia. The present study describes epidemiological, clinical, and pathological features of cases of bovine babesiosis diagnosed over the last two decades in the state of Mato Grosso do Sul in central-western Brazil. Examination of a total of 5298 bovine samples resulted in a definitive or presumptive diagnosis of babesiosis in 41 cases from 33 outbreaks. Babesia bovis was the most common causal agent of bovine babesiosis, and animals of all ages were affected. Zebu cattle, which are more frequent in the study region than are taurine cattle, were the cattle most often affected. Although zebu cattle have been claimed to be highly tolerant, the present findings show that they can develop severe illness and die due to Babesia spp. infection. The disease was diagnosed almost every year over the time period covered in this study (1995-2017). However, the number of cases of bovine babesiosis diagnosed was low compared to the total number of cases reviewed. Veterinary practitioners that work on cattle farms should to be able to diagnose the disease in its early stages so that treatment can be initiated promptly and economic losses can be minimized.

Clinical and therapeutic aspects of an outbreak of canine trypanosomiasis.

Trypanosomiasis caused by Trypanosoma evansi can seriously affect both domestic and wild animals. This article reports on an outbreak of canine trypanosomiasis on a farm in the Pantanal region of Brazil. The farm had 38 dogs, 20 of which died before receiving veterinary care. The remaining 18 dogs were underwent anamnesisn, clinical examination, hematological and biochemical evaluations. Blood smears and PCR analysis were performed for the diagnosis. The treatment protocols used according to the clinical recovery or parasitological cure of the dogs, using diminazene diaceturate, isometamidium chloride or quinapyramine sulfate. Post-treatment parasitological evaluation was performed by the microhematocrit technique. 7/18 dogs were PCR positive for T. evansi (confirmed by sequencing). There was clinical findings, which were consistent with both the acute and chronic stages of the disease in dogs. The infected dogs all exhibited at least one clinical sign of the disease. The hematological findings were compatible with trypanosomiasis, highlighting the hypochromic microcytic anemia as the main outcome. No treatment protocol was fully effective and the prolonged use of diminazene diaceturate caused the death of an animal. The trypanosomiasis can cause high rates of morbidity and mortality in dogs and difficulty in establishment an effective and safe therapeutic protocol.

Eimeria species in dairy and beef cattle of different ages in Goiás state, Brazil.

This study aimed to identify the frequency of occurrence of Eimeria species in Holstein (taurine dairy cattle) and Nelore (zebu beef cattle) on 12 farms in the state of Goiás, Brazil. A total of 2,601 animals were evaluated, and the cattle were divided into the following age-group categories: up to three months old, four to eight months old, nine to 16 months old and cows. Fecal samples were collected directly from the rectum of all animals. None of the cattle presented any clinical signs of eimeriosis during the survey. From the results obtained, it could be concluded that higher rates of infection by Eimeria spp. were observed in Holstein cattle between the 1st and 16 th months of age and in Nelore cattle between the 1st and 8th months of age. Twelve and nine Eimeria species were diagnosed in Holstein and Nelore cattle, respectively. However, Eimeria zuernii was more prevalent in animals up to three months old, while Eimeria bovis was more common in the remaining categories (cattle aged four months and over). Future studies on the epidemiology and/or seasonality of infection by Eimeria species in cattle, from birth onwards, are needed in order to better understand the dynamics of E. zuernii and E. bovis occurrences, especially in cattle under three months of age.

Comparison of McMaster and Mini-FLOTAC fecal egg counting techniques in cattle and horses.

The aim of this study was to compare two fecal egg count (FEC) techniques; McMaster (McM) and Mini-FLOTAC (mF), for the detection of cattle and horse gastrointestinal nematode eggs, in different locations. Experiment 1: feces were collected from 16 cattle and FEC was performed individually, using mF with the sensitivity of 5 eggs per gram of feces (EPG) and McM with a sensitivity of 50 EPG at Empresa de Pesquisa Agropecuária de Minas Gerais - EPAMIG and the Laboratory of Parasitic Diseases of the University of Parana - LDP/UFPR. Experiment 2: Fecal samples from 30 horses were analyzed with mF (sensitivity of 5 EPG) and McM (sensitivity of 25 EPG) at the University of Mato Grosso do Sul - UFMS and LPD/UFPR. Experiment 3: feces were collected from 14 foals and FEC was performed using mF (sensitivity of 5 EPG); and McM (sensitivity of 25 EPG) only at the LPD/UFPR. For cattle, the average FEC of mF was 962 at LPD; and 1248 at EPAMIG; for McM it was 1393 at LPD and 1563 at EPAMIG. For horses, the FEC average using the mF was 650 at LPD and 469 at UFMS; and for McM it was 677 at LPD and 554 at UFMS. For foals, the average FEC for mF was 404 and 436 for McM. In all experiments, the standard deviation and the coefficient of variation values were significantly lower for mF. Therefore, it is recommended the use of the Mini-FLOTAC technique, which is a method with less variability and higher accuracy, particularly for animals with low FEC.

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.

Multispecies resistance of cattle gastrointestinal nematodes to long-acting avermectin formulations in Mato Grosso do Sul.

The use of long-acting avermectins (AVMs) in cattle to treat infections with gastrointestinal nematodes was common in Brazil until its prohibition by state authorities. The prohibition; however, was rescinded in 2015, but a scientific discussion of the pros and cons of the use of these formulations is necessary. We evaluated the levels of resistance to 1.0 and 3.5% doramectin and to 3.15% ivermectin in cattle. The worms in animals treated with 3.5% doramectin were characterized by the suppression of oviposition and by a higher proportion of adult females carrying no eggs. Haemonchus placei, Cooperia punctata, C. pectinata, C. spatulata, and Oesophagostomum radiatum were resistant to the above compositions. The administration of long-acting AVM formulations did not result in a higher efficacy against these helminth populations.

First genetic identification of Cryptosporidium parvum subtype IIaA14G2R1in beef cattle in Brazil.

The presence of Cryptosporidium spp. in a cattle herd registered with an outbreak of diarrhea was investigated and the the molecular subtyping of Cryptosporidium parvum was characterized. Fecal samples from 85 Nellore beef cattle (Bos indicus) were collected and examined with Ziehl-Neelsen modified staining method. Fifty-four cattle (63.52%) had Cryptosporidium spp. oocysts in their feces. Fragments of genes encoding the 18S ribosomal RNA subunit and a 60-kDa glycoprotein (gp60) were amplified by nested PCR accomplished in the 11 most heavily parasitized samples, and the amplicons were sequenced. Eight of the 11 analyzed samples were positive for 18S rRNA sequences and identified monospecific infections with C. parvum. Seven samples were positive for gp60 and identified subtypes IIaA15G2R1 (6/11) and IIaA14G2R1 (1/11). This report is the first for C. parvum subtype IIaA14G2R1 in beef cattle in Brazil.

Clinical and therapeutic aspects of an outbreak of canine trypanosomiasis / Aspectos clínicos e terapêuticos de um surto de tripanossomíase canina

Abstract Trypanosomiasis caused by Trypanosoma evansi can seriously affect both domestic and wild animals. This article reports on an outbreak of canine trypanosomiasis on a farm in the Pantanal region of Brazil. The farm had 38 dogs, 20 of which died before receiving veterinary care. The remaining 18 dogs were underwent anamnesisn, clinical examination, hematological and biochemical evaluations. Blood smears and PCR analysis were performed for the diagnosis. The treatment protocols used according to the clinical recovery or parasitological cure of the dogs, using diminazene diaceturate, isometamidium chloride or quinapyramine sulfate. Post-treatment parasitological evaluation was performed by the microhematocrit technique. 7/18 dogs were PCR positive for T. evansi (confirmed by sequencing). There was clinical findings, which were consistent with both the acute and chronic stages of the disease in dogs. The infected dogs all exhibited at least one clinical sign of the disease. The hematological findings were compatible with trypanosomiasis, highlighting the hypochromic microcytic anemia as the main outcome. No treatment protocol was fully effective and the prolonged use of diminazene diaceturate caused the death of an animal. The trypanosomiasis can cause high rates of morbidity and mortality in dogs and difficulty in establishment an effective and safe therapeutic protocol.

Resumo A tripanossomíase causada por Trypanosoma evansi pode acometer gravemente os animais domésticos e selvagens. Este artigo relata um surto de tripanossomíase canina em uma fazenda na região do Pantanal, Brasil. Na fazenda havia 38 cães, 20 dos quais morreram antes de receber cuidados veterinários. Os 18 cães restantes foram submetidos a anamnese, exame clínico, avaliação hematológica e bioquímica. Esfregaços de sangue e análise da PCR foram realizados para o diagnóstico. Os protocolos de tratamento foram utilizados de acordo com a recuperação clínica ou cura parasitológica dos cães, utilizando diaceturato de diminazeno, cloreto de isometamídio ou sulfato de quinapiramina. A avaliação parasitológica pós-tratamento foi realizada pela técnica de microhematócrito. 7/18 cães foram PCR positivos para T. evansi (confirmado por sequenciamento). Os achados clínicos encontrados, foram consistentes com os estágios agudo e crônico da doença em cães. Todos os cães infectados exibiram pelo menos um sinal clínico da doença. Os achados hematológicos foram compatíveis com a tripanossomíase, destacando a anemia microcítica hipocrômica como principal consequência. Nenhum protocolo de tratamento foi totalmente eficaz e o uso prolongado de diaceturato de diminazeno causou a morte de um animal. A tripanossomíase pode causar altas taxas de morbidade e mortalidade em cães e dificultar o estabelecimento de um protocolo terapêutico eficaz e seguro.