Trypanosoma vivax in and outside cattle blood: Parasitological, molecular, and serological detection, reservoir tissues, histopathological lesions, and vertical transmission evaluation.

This study reports assessment of the sensitivity of diagnostic techniques to detect T. vivax in experimentally infected cattle. Additionally, it describes T. vivax extravascular parasitism during the acute and chronic phases of trypanosomosis and congenital transmission. The T. vivax diagnosis was compared using blood samples collected from the jugular, coccygeal and ear tip veins. For this study, 13 males and two females were infected with ≈ 1 × 106 viable T. vivax trypomastigotes (D0). One animal was kept as a negative control during the entire study. The 13 infected males were euthanized between 14 and 749 days post-infection (DPI). After confirming the cyclicity of both females (9 months of age), they were naturally mated with a bull. One female was euthanized at 840 DPI, and the other at 924 DPI. The two calves, one from each female, were euthanized at six months of age (924 DPI), and the negative control at 924 DPI. During this period, T. vivax in blood was assessed using direct methods (Woo test, cPCR, microscopic examination of fresh wet blood films and parasite quantification - Brener method), and serological methods (IFAT, ELISA, and IA). Tissue samples were collected from the liver, spleen, brain, cerebellum, heart, testicles, epididymis, kidneys, eyeballs, pre-scapular lymph nodes, ear tips, mammary glands, uterus, and ovaries. The protozoan DNA was examined using LAMP. There was no difference in the detection of T. vivax using the Woo test and Brener method among the jugular, coccygeal, and ear tip veins. The sensitivity of the detection methods varied depending on the disease phase. Direct methods (Woo test, Brener method, and cPCR) demonstrated higher sensitivity during the acute phase, while serological methods (IFAT, ELISA, and IA) were more sensitive during the chronic phase. Anti-T. vivax antibodies were detected up to 924 DPI. Tissue evaluation using LAMP demonstrated the presence of T. vivax DNA and associated histopathological changes up to 840 or 924 DPI. Only in mammary glands and ovaries was no DNA detected. The most frequently observed histopathological alteration was lymphohistioplasmocytic inflammatory infiltrate. No transplacental transmission of T. vivax was observed.

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.

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.

How many cattle can be infected by Trypanosoma vivax by reusing the same needle and syringe, and what is the viability time of this protozoan in injectable veterinary products?

It was investigated how many cattle become infected with Trypanosoma vivax by subcutaneous (SC), intramuscular (IM) and intravenous (IV) routes, using the same syringe and needle from an animal with acute T. vivax infection. Besides, the T. vivax viability in 109 injectable veterinary drugs (antibiotics, antiparasitics, reproductive hormones, vitamin complex and derivatives, vaccines, anaesthetics, anti-inflammatory/antipyretics, antitoxics). In the field assay, four groups were performed: T01, T02 and T03 animals that received saline solution with the same syringe and needle contaminated with T. vivax via SC, IM and IV routes, respectively, and T04 control animals that received only saline solution with the same syringe and needle IV. In the laboratory, drugs had their pH measured and T. vivax viability verified. The number of cattle infected with T. vivax via SC (3/20) was lower (P ≤ 0.05) compared to via IM (9/20), which was lower (P ≤ 0.05) compared to IV (15/20). The solution pH did not influence T. vivax viability. In 44% (48/109) of the products, T. vivax remained viable regardless of time, stooding out that in 100% of oxytocins the protozoan was verified, at some evaluation times. The mean of T. vivax quantified in foot-and-mouth and brucellosis vaccines and in doramectin-based products were higher (P ≤ 0.05) than found in blood + saline solution.

In vitro and in vivo effectiveness of disinfectants against Trypanosoma vivax.

Trypanosoma vivax causes bovine trypanosomosis in cattle and resulting in economic losses to farmers. In Brazil, shared contaminated materials is the main transmission pathway. To evaluate the effectiveness of different disinfectants for T. vivax, in vitro and in vivo analyses were performed. At the laboratory, 21 disinfectants were tested. The disinfectants were placed in microtubes containing blood with approximately 1.0 × 106 trypomastigotes of T. vivax. The viability and motile of trypomastigotes after 30 s, one, 10, 15 and 30 min was evaluated by the thick drop method and the efficacy calculated. Disinfectants that showed 100% effectiveness were used in in vivo tests. Thirty calves negative for T. vivax were divided into six groups and were inoculated with disinfectant solutions (46% alcohol, 70% alcohol, or 0.5% iodine) + 1 × 106 trypomastigotes of the protozoa. Blood from each animal was collected at seven, 14 and 21 days after inoculation to verify the viability and presence of the protozoan by Woo, Brener, PCR, and LAMP methods. In the in vitro step, 13 of the 21 disinfected solutions exhibited 100% effectiveness against T. vivax at all evaluation times. In contrast, 70% alcohol and 0.5% iodine solutions exhibited 100% effectiveness in the in vivo tests and can be used to disinfect needles and syringes. The use of disinfectants is a rapid and efficient procedure to disinfect materials utilized in the field and concomitantly could help to reduce the dissemination of T. vivax in the cattle herd in cases of iatrogenic transmission.

Eimeria spp. in naturally infected beef cattle: Dynamics of oocysts excretion, prevalence, and comparison between parasitological diagnostics.

Eimeria spp. infections cause mortality, reduced well-being, and substantial economic losses implications for cattle production worldwide. The present work followed up the excretion of Eimeria spp. oocysts in two naturally infected beef herds, from two different properties, to investigate the dynamics of oocyst excretion and the prevalence of Eimeria spp. in different animal categories and seasons of the year (rainy season - October to April; dry season - May to September). Even that, the species of Eimeria were identified and the parasitological techniques of Gordon and Whitlock modified and Mini-FLOTAC were used. In both herds, animals up to 14 months had a mean total OPG counts higher than older animals (after 15-16 months of age), and the species E. zuernii and E. bovis were more frequently identified, the first species being more frequent in animals from 1 to 2 months of age, while E. bovis prevailed from three months old. On property 1, the highest mean OPG counts (P ≤ 0.05) were obtained between October 2017 and September 2018, with the highest mean OPG counts in October 2017, when the animals were aged between 4-5 months. The prevalence of the pathogen on property 1 was 59.16 % and 43.62 % in the rainy and dry season, respectively, a higher parasitic load (P ≤ 0.05) was verified in the rainy season. On property 2, the mean OPG counts of Eimeria spp. was higher (P ≤ 0.05) in animals between 8-16 months, with the highest peak in November 2019, when they were one year old. The on-site prevalence during the rainy season on property 2 was 53.09 % and 49.79 % on dry season, and no difference (P = 0.92) in the mean OPG counts of Eimeria spp. during the seasons. There was a difference (P ≤ 0.05) in the count of oocysts in females after 18 months of age than males, which was probably due to the increase in animal density. Both tested techniques can be used for quantification of the excretion of oocysts of Eimeria spp. in cattle feces showing the same OPG mean count (r = 0.9287; p = 0.0025; R² = 0.8625). Mini-FLOTAC showed higher prevalence for Eimeria spp., however, can be an obstacle depending on the number of fecal samples that need to be processed.

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.

Evaluation of rotational grazing as a control strategy for Rhipicephalus microplus in a tropical region.

Rhipicephalus microplus is the most significant tick of livestock and its control is particularly challenging due to its resistance to commercial acaricides. Pasture rotation is considered a management strategy that could help control R. microplus, however, the literature only contemplates mathematical models and little is known about the effects of this practice in the field. The objective of this work was to determine whether pasture rotation is an efficient method for controlling R. microplus. Two different experiments were performed that involved groups of continuous and rotational grazing bovines. Female ticks measuring 4.5-8.0 mm were counted on animals while larvae in pasture were counted using the flannel drag technique. Treatment for infested bovines was applied when the average group tick count was ≥30 females. The results showed that rotational grazing (with 20-day periods of rest) had a higher tick count on-host than continuous grazing (P < 0.05) and additional bovine treatment was needed. Sixty and 105 days were needed to re-infest and disinfest pasture of R. microplus larvae, respectively. The first treatment of bovines occurred 91 days after the animals were placed in a closed area. The results indicate that rotational grazing is not an efficient way to control R. microplus.

Do rainfall and tick burden affect the efficacy of pour-on formulations against Rhipicephalus (Boophilus) microplus?

Rhipicephalus (Boophilus) microplus is the most important livestock tick, causing economical losses especially in tropical and subtropical regions. Pour-on formulations using synthetic chemicals, remain the most farmer friendly conventional form of tick control method. Misuse of acaricides can lead to the emergence of resistance, residual chemicals in animal products, the poisoning of vertebrates and environment contamination. Despite the increase in the use of pour-on formulations, little is known regarding the therapeutic and residual efficacy of these products after the treated animals are exposed to rainfall. Moreover, information is scanty on whether efficacy is modulated by different levels of tick burden. Three studies were conducted. In the two first experiments, we evaluated the therapeutic and residual efficacies of commercial pour-on products (fluazuron 2.5 mg/kg and fipronil 1.0 mg/kg, respectively) on cattle naturally infested with R. (B.) microplus under the condition of simulated rainfall. In the third study, we investigated whether tick burden affects the efficacy of the drugs used a chemical control method involving three different formulations (Day 0: cypermethrin 5.0 mg/kg + chlorpyrifos 7.0 mg/kg + citronellal 0.5 mg/kg; Day 7: fluazuron 3.0 mg/kg + abamectin 0.5 mg/kg and Day 56: fipronil 1.0 mg/kg + fluazuron 3.0 mg/kg). Tick counts were performed on different days to assess efficacy. It was observed that the residual efficacy of the commercial formulations was lower on animals that received simulated rain. In addition, therapeutic and residual efficacies of the products were reduced in the group of animals with a high tick burden of R. (B.) microplus. In conclusion, rainfall decreases the efficacy of acaricides. Also, the tick burden must be considered when selecting a pour-on formulation.