Evaluation of mechanical transmission of Trypanosoma vivax by Stomoxys calcitrans in a region without a cyclic vector.

This work investigated the mechanical transmission of Trypanosoma vivax by Stomoxys calcitrans to cattle in a region without a cyclic vector. The study involved two experiments, one with calves experimentally infected with T. vivax, in the acute phase of trypanosomosis (Experiment 1) and the other in the chronic phase (Experiment 2). In both experiments, two transmission methods were used with flies that had not fed for 24 h or had never fed: (i) Method 1: flies released freely in cattle pens (≈3,300 flies/pen for 10 days); and (ii) Method 2: flies placed in a feeding chamber (12 flies/animal). To develop Method 1 in the two experiments (acute and chronic phases), T. vivax-positive animals were kept with T. vivax-negative animals. Periodically, the Brener method, Woo method, blood smears, cPCR, ELISA, IFAT, and Imunoteste® were performed to detect T. vivax in the animals. We also recorded the animals' head tossing and hoof stomping and the number of flies near the pens' inner walls. Subsequently, biological testing was performed using lambs. For Method 2 in both experiments, flies inside the feeding chamber first fed on T. vivax-positive animals and later on negative animals. In both experiments and methods, we examined the flies for the presence of T. vivax through blood smears and cPCR of the proboscis and abdomen. In Experiment 2 (chronic phase), a test was conducted to determine how long trypomastigotes forms could survive on the blood of animals with different levels of parasitemia. None of the animals (calves and lambs) became infected with T. vivax or showed antibodies against it. During the evaluation period, the animals in the presence of the flies exhibited more hoof stomping and head tossing compared to those without flies (control). Additionally, there was an increase in the number of flies in the pens during the experiment. Only in Experiment 1 (acute phase) were T. vivax trypomastigotes and DNA found in the abdomen of the flies but not in the proboscis. In Experiment 2 (chronic phase), higher concentrations of trypomastigotes per milliliter of blood were associated with a shorter the lifespan of this stage of the parasite. In conclusion, under the variable conditions of the experiments (hosts, number of flies, and level of parasitemia), S. calcitrans was unable to mechanically transmit T. vivax to cattle.

First report of the efficacy of a fluralaner-based pour-on product (Exzolt® 5%) against ectoparasites infesting cattle in Brazil.

BACKGROUND: This study describes the effectiveness of a novel active pharmaceutical ingredient, fluralaner (isoxazoline class), against important ectoparasites infesting cattle in Brazil. METHODS: A total of 13 studies involving a 5% fluralaner-based pour-on formulation (Exzolt 5%; further referred to as Exzolt) were conducted. Specifically, the effectiveness of this formulation was studied against Rhipicephalus microplus (6 studies), Cochliomyia hominivorax larvae (4 studies), Dermatobia hominis larvae (1 study) and Haematobia irritans flies (2 studies). RESULTS: The therapeutic efficacy of Exzolt was found to exceed 98% at 4 days post treatment (DPT), while persistent efficacy (> 90% efficacy) against repeated infestations of R. microplus was observed for up to 79 DPT. In field studies, ≥ 98% therapeutic efficacy was demonstrated at all study sites by 7 DPT, and a persistent efficacy (> 90% efficacy) was observed for 42, 49 or 56 DPT. Exzolt prevented C. hominivorax eggs from developing to the larval stage, thus mitigating the development of myiasis in cattle naturally and artificially infested with this screworm. The efficacy of Exzolt against D. hominis larvae was 98% at 3 DPT, while persistent efficacy (> 90% effectiveness) was found to last for up to 70 DPT. Against H. irritans, Exzolt showed therapeutic efficacy (≥ 90%) within the first day of treatment at both study sites, while persistent efficacy (≥ 90%) was observed for 7 DPT at one site and for 21 DPT at the other site. CONCLUSIONS: Overall, the results from these studies confirm that Exzolt is therapeutically efficacious against the most important ectoparasites infesting cattle in Brazil. The novel active pharmaceutical ingredient, fluralaner, provides a new treatment option for farmers to control cattle ectoparasites, especially where there is resistance to other chemical classes. In addition, an effective control of ectoparasites will improve overall cattle health and well-being as well as production.

FIRST REPORT OF GENUS Cryptosporidium IN CERVIDS SPECIES: Mazama americana, Mazama nana AND Blastocerus dichotomus.

We analyzed Cryptosporidium spp. in fecal samples of wild cervids (Ozotoceros bezoarticus, Blastocerus dichotomus, Mazama nana, Mazama americana, and Mazama bororo) from many Brazilian regions, a fact unprecedented in the literature. Sniffer dogs were used to collect 936 fecal samples of cervids from 14 Brazilian localities. Cervids species were identified using polymerase chain reaction (PCR) performed from genomic DNA extracted from 563 fecal samples of Ozotoceros bezoarticus, Blastocerus dichotomus, Mazama nana, Mazama americana, and Mazama bororo. Cryptosporidium spp. oocyst screening was performed using malachite green negative staining. Nested PCR (nPCR) protocols targeting the 18S rRNA and GP60 genes followed by genetic sequencing were performed for Cryptosporidium spp. detection and Cryptosporidium parvum subtyping, respectively. Nested PCR targeting actin gene and genetic sequencing were performed in samples with non-identified Cryptosporidium species by 18S rRNA amplicon sequencing. The association between the occurrence of Cryptosporidium and the presence of bovines in the same locality was evaluated using Fisher's exact test. The positivity rates of diagnostic methods were compared by McNemar test and the Kappa correlation coefficient. The prevalence rates of Cryptosporidium spp. in cervids were 1.42% (8/563) and 0.36% (2/563) by nPCR and malachite green negative staining, respectively. C. parvum IIaA16G3R1 isolate was identified in three fecal samples from M. americana, two from M. nana and one from B. dichotomus. Cryptosporidium ryanae were found in one sample from B. dichotomus. We identified a new Cryptosporidium genotype, named Cryptosporidium deer genotype BR, from one M. americana fecal sample.

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.

Viability of Haemonchus placei parasitism in experimentally infected young goats.

The present study aimed to evaluate the viability of Haemonchus placei parasitism in experimentally infected goats. For that, 14 75 days old kids male Saanen kids were placed in one of the four experimental groups: GI - infected with 5000 H. placei L3 (n = 4); GII - infected with 5000 H. contortus L3 (n = 4); GIII - infected with 2500 H. contortus L3 + 2500 H. placei L3 (n = 4), and GIV - control, inoculated with distilled water (n = 2). Each kid received, orally, the infective dose in a single inoculum. Based on daily fecal egg counts, the average pre-patent period was determined as 24 days for H. contortus, and 31 days for H. placei. Regarding the Haemonchus spp. recovered at necropsy, the experimental groups GI, GII, and GIII had, respectively, an average of 25.5, 619.5, and 724.75 (120 H. placei, and 604.75 H. contortus) adult specimens, and no immature forms. Under the conditions of this study, the viability of goat infection by H. placei was confirmed, although, with low susceptibility. Nevertheless, the parasitism of this helminth species was more intense when associated with H. contortus. This fact indicates that in common grazing between cattle and young goats, when the latter end up ingesting both Haemonhcus species, especially in a mixed infection, H. placei may also parasitize them.

Spatial distribution and risk factors for Toxoplasma gondii seropositivity in cattle slaughtered for human consumption in Rondônia, North region, Brazil.

The present study aimed to evaluate Toxoplasma gondii seroprevalence in cattle slaughtered for human consumption from rural properties in the state of Rondônia, North region, Brazil; the seroprevalence was determined using indirect immunofluorescence assays (IFATs). Additionally, spatial distribution and risk factors associated with toxoplasmosis were also analyzed. Of the 1000 cattle serum samples examined, 53 (5.3%) were determined to be seropositive for T. gondii with antibody titers (IgG) ≥64. In regard to results of the studied risk factors (presence of cats, cats with free access to cattle, breeding system, animal's gender, consumption of raw milk by humans on the property and cattle abortion in the last 12 months) and the odds ratio (OR) of each of these factors influencing cattle to acquire toxoplasmosis, only animals raised on a feeder/stocker/backgrounder system presented a higher probability of being seropositive for T. gondii (OR≥1, P=0.04) than cattle raised only in a feeder/stocker system. There was no association between the occurrence of reproductive problems and T. gondii seropositivity. Based on results obtained in the Brazilian state of Rondônia, it could be concluded that the presence of cats and their contact with cattle on each property, cattle breeding purpose and cattle abortion in the last 12 months were not considered risk factors for T. gondii infection in cattle. Considering that the presence of T. gondii was detected in animals slaughtered in the state of Rondônia, consuming raw or undercooked meat from seropositive cattle should be considered a route of transmission of T. gondii to humans. However, the prevalence of toxoplasmosis diagnosed in cattle from this state (5.30%) is lower than the prevalence of toxoplasmosis observed in South, Southeast and Center-West regions of Brazil, which may vary between 48.5% and 71.0%. The low prevalence of toxoplasmosis in cattle is highlighted in Rondônia, which is the sixth largest state for cattle slaughtering in Brazil and is responsible for producing 20% of all cattle meat exported by Brazil.