Strategic control of the cattle tick Rhipicephlaus microplus applied to rotational and silvopastoral grazing systems in subtropical areas.

This work evaluated if strategic control based on no more than three or four annual treatments is useful to control Rhipicephalus microplus infestations on cattle when it is applied to intensive rotational grazing and silvopastoral systems with high stocking rates in subtropical areas. In the intensive rotational grazing system, three annual treatments with chemical acaricides were applied on cattle in two different schemes: between spring and early summer and from late winter and late spring. Strategic control based on three treatments with chemical acaricides from late winter to late spring plus an additional fourth treatment in summer was tested in the silvopastoral system. In the intensive rotational grazing systems, the control schemes allow to reach a significant reduction in the tick load on cattle considering a time interval from spring to autumn. However, the efficacy levels were not high enough in some specific moments, namely, the tick counts of summer and autumn (there were not significant differences between treated and control groups). The scheme evaluated in the silvopastoral grazing system yielded better results than those tested for the intensive rotational system, because significant differences in tick load between treated and control groups were observed in all post-treatment counts and when the analysis was performed for the whole study period. However, values of efficacy in the count-by-count comparison were disparate, ranging from 64.1 to 99.7. Although the efficacy values obtained in the silvopastoral system were better than those of the rotational grazing systems, the total tick load on treated cattle in autumn was not low enough (mean abundance values 25.14 and 38.14). Ticks were more evenly distributed among hosts in late summer and autumn than in spring or early summer, where few hosts carry most of the ticks. Some management strategies as intensive rotational systems or silvopastoral structures can lead to a more efficient forage use, but they imply greater tick challenge than in extensive grazing systems. In these situations, the schemes of strategic control bases on three or four annual treatments should be complemented with additional tactical treatments in late summer or autumn.

Comparative analysis of Rhipicephalus microplus (Canestrini, 1888) infestation in pure breed (Hereford) and cross breed (Braford) cattle herds subjected to the same chemical treatments.

The aim of this work was to analyze the R. microplus (Canestrini, 1888) infestation in two bovine herds with different degrees of natural resistance (i.e., Hereford and Braford) to ticks subjected to an identical chemical treatment scheme to ticks at the same farm, to demonstrate the impact on tick control of the incorporation of a more resistant bovine breed. Two groups of ten Hereford and Braford cows each were subjected to eleven chemical treatments between August 2022 and October 2023 (four fluazuron, two fipronil 1%, one ivermectin 3.15% and four immersion in a dipping vat with a combination of cypermethrin 10% and ethion 40%). Tick population was shown to be susceptible to ivermectin, fluazuron and the mix cypermethrin 10%-ethion 40% and resistant to fipronil according to in vitro tests. Tick infestation was significantly greater in the Hereford cows than in the Braford cows. Tick infestation in both Hereford and Braford breeds was similar when treatment with functional drugs was applied, but when a block of the treatments was done with drugs with decreased functionality due to resistance (i.e. fipronil), treatment failure was manifested more strongly in the most susceptible breed. The incorporation of cattle breeds with moderate or high resistance to R. microplus is instrumental to optimize the efficacy and sustainability of chemical control of ticks in a scenario where resistance to one or more chemical groups is almost ubiquitous, because it favors the biological control of this parasite.

Does parity order or cow antibody level influence the humoral immunity of calves for cattle anaplasmosis?

Anaplasmosis is a widely distributed vector-borne disease of cattle caused by the bacteria Anaplasma marginale, which may lead to severe losses in beef and dairy cattle production. Anecdotal information among farmers suggested that some cows may deliver calves more resilient to anaplasmosis. To investigate this, we tested two hypotheses: (i) whether the parity order or (ii) cow antibody levels may influence the humoral immunity of pre-weaning calves against cattle anaplasmosis. For this study, we sampled 170 cattle (Bos taurus taurus, Angus breed) on a farm in Southern Brazil, comprising 85 cows (50 multiparous and 35 primiparous) and their 85 calves (50 days old). Antibodies against A. marginale were investigated using iELISA. Eighty percent of the animals were considered seropositive (100% of the cows and 60% of the calves). There was no significant difference in seroprevalence between calves from primiparous and multiparous cows. However, calves from multiparous cows exhibited higher antibody levels. To address the second question, we classified cows based on their antibody levels to A. marginale (high or low). Calves from cows with high antibody levels also showed elevated antibody levels against A. marginale. Furthermore, calves from cows with high antibody levels had approximately four times greater odds of being seropositive for A. marginale at 50 days old than those born to mothers with low antibody levels. In conclusion, the calf's antibody level against A. marginale appears to be related to the cow's parity order and the mother's antibody level.

Resistance of the cattle tick Rhipicephalus (Boophilus) microplus to fluazuron in Argentina.

The aim of this work is to report the presence of resistance to fluazuron in a population of Rhipicephalus microplus in Argentina. The evidence was obtained from field and in vitro trials. In the field trial, cattle infested with ticks was treated with two commercial formulations of fluazuron. The in vitro trial (adult immersion test, AIT) was performed by using technical grade fluazuron. In the field trial, there were no significant differences between the treated and control groups between days 2 and 34 post-treatment. The only exceptions (treated group I in day 14 post-treatment, treated group II in days 23 and 29 post-treatment) had a significantly lower tick load than the untreated group, but the efficacy was not higher than 70%. Viable engorged females were collected on both groups of treated bovines in all counts, and the production of viable larvae was not precluded with the application of the two commercial formulations of fluazuron evaluated in this study. The results obtained with the in vitro assay (AIT) also indicate that the R. microplus population tested in this work has a higher level of resistance to fluazuron than another susceptible field strain. The integrated analysis of the field and in vitro trials clearly reveals the emergence of resistance to fluazuron in a R. microplus population from Argentina. This diagnosis of resistance does not imply that the fluazuron has lost its functionality at a regional scale, but it highlights the need to establish control strategies that minimize the use of this drug in order to preserve its functionality as an acaricide.

Alternative applications of the strategic control against the cattle tick Rhipicephalus (Boophilus) microplus in a subtropical area.

Although different evaluations on the efficacy of the strategic control against Rhipicephalus microplus have been performed, the effects of successive applications of these schemes on the abundance of cattle ticks have not been evaluated. The aim of this work was to analyse the long-term effect of strategic applications of chemical acaricides on the R. microplus infestation in cattle in a subtropical area. These schemes are based on the application of three annual treatments between late winter and late spring. Additionally, a trial to evaluate the efficacy of the strategic control by deferring the first treatment from late winter to spring and the third treatment from late spring to summer was also carried out. The efficacy of the strategic control applied on 3 consecutive years was significant. The tick infestation in the treated group always remained at low levels, because mean number of ticks was almost never higher than 20. Regarding the trial where the third application of acaricide was deferred from spring to summer, and the first one from late winter to spring, the differences between treated and control group were significant in all post-treatment counts. The results of this study add evidence that support the sustainability of the strategic control in subtropical areas where the population dynamics of R. microplus is characterized by a well-marked seasonal pattern. Three relevant aspects were determined: (i) the feasibility and efficacy of successive applications of the strategic control in consecutive years; (ii) the time window to start the sequence of treatments is from late winter to mid-spring; (iii) it is achievable deferring the last treatment from late spring to summer if the tick infestation levels on cattle are low enough to allow it.

In vitro diagnosis of resistance of the cattle tick Rhipicephalus (Boophilus) microplus to fipronil in Argentina.

The aim of this work was to adapt and apply the in vitro larval bioassay packet test (LPT) to diagnose resistance of Rhipicephalus microplus ticks to fipronil in Argentina. Ten serial dilutions of fipronil were included in the LPT bioassay. Seven populations of R. microplus from six Argentinean Provinces were evaluated. Samples from Colonia Benítez, Quimilí and Intiyaco were unambiguously classified in the category 'resistant'. In all these cases the 95% confidence intervals did not overlap with that of the susceptible reference strain and the slope values were lower than the slope value obtained for the susceptible reference strain. The resistance ratio (RR50) values of the populations from Colonia Benítez I and II, Quimilí and Intiyaco were 2.23, 3.49, 6.84 and 3.39, respectively. Populations from El Colorado I and II, Acambuco, Vera and Feliciano were classified as 'susceptible' because the 95% confidence intervals overlapped with that of the susceptible reference strain and the RR50 values never exceeded 1.32. The results of these LPT bioassays indicate the presence of fipronil resistant R. microplus populations in Argentina.

The membrane domain of vacuolar H(+)ATPase: a crucial player in neurotransmitter exocytotic release.

V-ATPases are multimeric enzymes made of two sectors, a V1 catalytic domain and a V0 membrane domain. They accumulate protons in various intracellular organelles. Acidification of synaptic vesicles by V-ATPase energizes the accumulation of neurotransmitters in these storage organelles and is therefore required for efficient synaptic transmission. In addition to this well-accepted role, functional studies have unraveled additional hidden roles of V0 in neurotransmitter exocytosis that are independent of the transport of protons. V0 interacts with SNAREs and calmodulin, and perturbing these interactions affects neurotransmitter release. Here, we discuss these data in relation with previous results obtained in reconstituted membranes and on yeast vacuole fusion. We propose that V0 could be a sensor of intra-vesicular pH that controls the exocytotic machinery, probably regulating SNARE complex assembly during the synaptic vesicle priming step, and that, during the membrane fusion step, V0 might favor lipid mixing and fusion pore stability.

Stochastic modelling to predict a priori the potential outcomes of different control schemes against the cattle tick Rhipicephalus microplus.

Stochastic models are valuable tools to describe and analyze tick population dynamic in a given area, and to evaluate different control schemes. The objective of this study was to provide a tool to predict a priori how a control scheme could affect the abundance of Rhipicephalus microplus in an area highly favourable for its development, through the building of a stochastic model of the population dynamic of this tick. The dynamic was stochastically modelled using field data of the parasitic and non-parasitic phases of R. microplus. The host susceptibility to tick infestations was also considered by including representative values of natural resistance of three different breeds (Bos indicus, B. taurus and B. indicus x B. taurus). Two different control schemes (strategic and threshold methods) using three annual applications of synthetic acaricides in different moments were evaluated. Furthermore, we have evaluated the impact of different moments of pasture spelling as a tool for tick control on the dynamic of the non-parasitic phases of R. microplus. The results showed that the model appropriately fits to field data and can simulate the annual variability in female ticks count in animals from different ecological characteristics. The model has also captured the variability that host susceptibility has on tick abundance. This fact was expressed in the different levels of tick abundance obtained for the three breeds of bovines. According to the model, strategic control applications of chemical acaricides have more efficacy than the threshold control method. The model indicates that a pasture spelling started in early or mid-summer generates a higher reduction in the number of available eggs in pastures. Analysis of the results allows to conclude that the model developed in this study was useful to predict a priori the performance of different control methods based on the strategic application of chemical acaricides or by applying pasture spelling.

Analysis of management factors influencing Anaplasma marginale transmission.

The aim of this study was to identify factors that play a key role in the epidemiology of bovine anaplasmosis by adapting a model primarily developed for cattle babesiosis. A cross-sectional observational study was conducted to study the proportion of calf herds in endemic stability/instability for A. marginale in a semi-arid area of Argentina. The A. marginale inoculation rate (h) was calculated from age-specific seroprevalence using double-antigen sandwich ELISA in 58 herds of 4.5-8.5-month-old calves. Herds were considered to be in endemic instability (EI) at h < 0.005 and, therefore, at risk of anaplasmosis outbreaks. A generalized linear model was performed to explore husbandry practices associated with differences in A. marginale transmission. Additionally, spatial clustering of herds with the same immunological status was analyzed using spatial scan statistics (SatScan, Bernoulli model). Spearman's correlation was used to explore a possible association between A. marginale h and Babesia bovis and B. bigemina h (data obtained in previous works). Almost half (43 %) of the herds were in the EI zone for A. marginale. Calves raised under forage combinations had a greater risk of being in EI (OR = 5.41, CI95 %OR = 1.43-20.41) than those reared exclusively on permanent pastures, where cattle density is higher (P = 0.01). Moreover, calves from herds treated only with pyrethroids to control ticks had more chances of being in EI (OR = 4.16, CI95 %OR = 1.12-15.38) than calves from herds receiving different acaricide combinations (P = 0.03). Calves from herds subjected to more than two treatments against Haematobia irritans had higher odds for EI (OR = 5.69, CI95 %OR = 1.24-26.11) than those from herds using fewer than two treatments (P = 0.02). The spatial analysis revealed no spatial clustering of the immune status of the herds (P = 0.67 and P = 0.74 for low and high incidence rates, respectively). A significant variation between farms was observed in A. marginale h (CV = 90.38 %). The correlation analysis revealed a strong epidemiological link of A. marginale h with B. bovis h (Rho=0.794, P<0.001) and B. bigemina h (Rho=0.839, P<0.001). Given that R. microplus is the only vector of B. bovis and B. bigemina in the region, the results of this work strongly suggest an active and significant role of R. microplus in the transmission of A. marginale.

Can herd seroprevalence be used as an indicator of enzootic stability for bovine anaplasmosis? Insights from a case-control field study in Brazil.

Bovine anaplasmosis presents a significant challenge to livestock production in tropical, subtropical, and temperate regions. For many years, the concept of enzootic stability/instability (initially established for babesiosis) and herd seroprevalence as an indicator of outbreak risks have been applied to anaplasmosis. However, this model has never been definitively validated for Anaplasma marginale. The objective of this study was to examine the relationship between herd immunity (seroprevalence) and the occurrence of anaplasmosis outbreaks in Southern Brazil. A case-control study was conducted, categorizing farms into two groups: cases (farms with a history of clinical anaplasmosis) and controls (those without anaplasmosis). Thirteen farms were identified as "cases", while 23 were identified as "controls". A substantial difference in seroprevalence distribution between the two groups was observed. The majority of "control" farms exhibited over 75% of animals with antibodies to A. marginale in both calves and heifers, whereas the majority of "case" farms had a seropositive cattle percentage below 75%. Additionally, twelve months after cattle serology tests, we conducted a prospective follow-up survey to identify any clinical cases of anaplasmosis. Statistical associations (P < 0.05) were found between both retrospective and prospective anaplasmosis outbreaks and the hypothetical threshold of herd seroprevalence (75%). We hypothesize that herd seroprevalence may be an indicator of the risk of occurrence of clinical anaplasmosis. It appears that the epidemiology of cattle anaplasmosis, at least in our conditions, aligns with the well-known model of enzootic stability/instability originally applied to bovine babesiosis.

Prothrombotic changes in diabetes mellitus.

Although our understanding of vascular pathology has greatly improved in recent years, the cellular and molecular mechanisms underlying the enhanced thrombotic propensity in type 2 diabetes mellitus (T2DM) remain incompletely characterized. Detrimental interactions between activated vascular cells (i.e., platelets, leukocytes, endothelial cells) and the vulnerable atheromatous plaque are a major determinant of the increased atherothrombotic burden in T2DM patients. Endothelial damage and accelerated senescence, impairment of the endothelial progenitor cell repair system, plaque neovascularization and inflammation, decreased clearance of detrimental molecules within the plaque, and increased expression of matrix metalloproteinases may collectively contribute to intraplaque hemorrhage and subsequent rupture. Notably, recent data demonstrates the central importance of the tissue factor-microparticle-mediated pathway in diabetic thrombophilia and cardiovascular complications. Acting as detrimental amplifiers of various biological responses (including thrombogenicity and plaque remodeling), microparticles have also emerged as a key marker of global vascular damage in T2DM patients. Available evidence suggests that targeting the tissue factor-microparticle pathway may be a promising approach for reducing the burden of the atherosclerotic complications of diabetes.

The enhanced cyan fluorescent protein: a sensitive pH sensor for fluorescence lifetime imaging.

pH is an important parameter that affects many functions of live cells, from protein structure or function to several crucial steps of their metabolism. Genetically encoded pH sensors based on pH-sensitive fluorescent proteins have been developed and used to monitor the pH of intracellular compartments. The quantitative analysis of pH variations can be performed either by ratiometric or fluorescence lifetime detection. However, most available genetically encoded pH sensors are based on green and yellow fluorescent proteins and are not compatible with multicolor approaches. Taking advantage of the strong pH sensitivity of enhanced cyan fluorescent protein (ECFP), we demonstrate here its suitability as a sensitive pH sensor using fluorescence lifetime imaging. The intracellular ECFP lifetime undergoes large changes (32 %) in the pH 5 to pH 7 range, which allows accurate pH measurements to better than 0.2 pH units. By fusion of ECFP with the granular chromogranin A, we successfully measured the pH in secretory granules of PC12 cells, and we performed a kinetic analysis of intragranular pH variations in living cells exposed to ammonium chloride.

A comparison of two in vitro bioassays to detect resistance of the cattle tick Rhipicephalus microplus to fipronil.

The aim of this work is to compare the sensitivity of two in vitro bioassays to detect resistant to fipronil in Argentinean populations of the cattle tick Rhipicephalus microplus. Two different larval bioassays prepared with technical grade (97%) fipronil were compared: larval immersion test (LIT) and larval packet test (LPT). Seven strains from different provinces were treated with both assays. Colonia Tabay, Colonia Benítez, Intiyaco and Quimili strains were considered resistant in both LIT and LPT bioassays. The 95% confidence intervals (IC95) for lethal concentration 50% (LC50) did not overlap with the susceptible reference strain (SRS) and all the values of RR50 obtained were higher than 2. Garabato and Federal strains were considered as susceptible for both techniques because the IC95 for the LC50 overlapped with those of the SRS and the RR50 values were lesser than 2. An ambiguous situation occurs with Reconquista strain. This strain was considered as susceptible with LPT and with incipient resistant after LIT trial. The analysis of the results indicates that both LIT and LPT trials have enough sensibility to differentiate resistant and susceptible strains, but LIT was more sensitive than LPT when the resistance is incipient.

Evaluation of the impact of successive acaricide treatments on resistance evolution in Rhipicephalus microplus populations: Monodrugs versus drug combinations.

The aim of this work was to comparatively evaluate the evolution of resistance in Rhipicephalus microplus tick populations exposed to successive treatments with monodrug-based formulations and combinations of them in the same commercial formulation. Thirty-six heifers, naturally infested with R. microplus, were divided into three groups (G) and subjected to three successive treatments, on days 0 (Nov-2021), 43 (Jan-2022) and 78 (Feb-2022), with the following formulations: I) ivermectin 3.15% (Ivomec Gold®) (GI), II) fipronil 1% (Ectoline®) (GII) and III) a combination of fipronil 2% and ivermectin 1% (Vaquero®) (GIII). From Nov-2021 to Dec-2022, counts of R. microplus were periodically performed to evaluate the tick infestation. Engorged females were collected at the beginning and end of the trial to determine the evolution of tick resistance to ivermectin and fipronil by in vitro bioassays. At the end of trial, GII and GIII had higher tick counts (39.18 ± 11.88 and 38.33 ± 14.31, respectively) than group I (5.11 ± 6.24) (P<0.05). The in vitro results shows that the resistance ratio (RR50) values after the treatments were higher for fipronil (5.584 and 5.649 for GII and GIII, respectively) than for ivermectin (1.165 and 1.088 for GI and GIII, respectively). In the group treated with the combination (GIII), the RR50 increased for both drugs simultaneously. These results suggest that the successive use of drug combinations could exacerbate the problem of multi-resistance of R. microplus to chemical acaricides.

The renal v-ATPase a4 subunit is expressed in specific subtypes of human gliomas.

Vacuolar H(+) -ATPases (v-ATPases) are multimeric proton pumps which acidify various intra-cellular organelles and may participate in pHe and pHi regulation in cancer cell lines. The ATP6V0A4 gene encodes the a4 subunit which is expressed in kidney and epididymis. Because we found a4 mRNA highly expressed in C6Bu1 glioma cell line, we measured it in 205 glioma biopsies and 11 brain biopsies from epileptic patients. a4 was absent in epileptic brain biopsies, but was expressed by 34% (11/32) of grade III oligodendrogliomas, independently of the chromosome 1p19q codeletion. a4 expression in grade III oligodendrogliomas and oligoastrocytomas without the 1p19q codeletion tended to be associated with a shorter overall survival of patients. We also observed a4 expression in biopsies of pilocytic astrocytomas (68%; 19/28) and gangliogliomas (37%; 6/16). In pilocytic astrocytomas a4 expression was associated with a tandem duplication of the 7q34 chromosome region, distant 0.5 Mb to the ATP6V0A4 gene locus. In conclusion, a4 expression identifies subtypes of oligodendrogliomas, pilocytic astrocytomas and gangliogliomas and may contribute to refine characterization of these tumors. © 2012 Wiley Periodicals, Inc.

Evaluation of a new pocket echoscopic device for focused cardiac ultrasonography in an emergency setting.

INTRODUCTION: In the emergency setting, focused cardiac ultrasound has become a fundamental tool for diagnostic, initial emergency treatment and triage decisions. A new ultra-miniaturized pocket ultrasound device (PUD) may be suited to this specific setting. Therefore, we aimed to compare the diagnostic ability of an ultra-miniaturized ultrasound device (Vscan™, GE Healthcare, Wauwatosa, WI) and of a conventional high-quality echocardiography system (Vivid S5™, GE Healthcare) for a cardiac focused ultrasonography in patients admitted to the emergency department. METHODS: During 4 months, patients admitted to our emergency department and requiring transthoracic echocardiography (TTE) were included in this single-center, prospective and observational study. Patients underwent TTE using a PUD and a conventional echocardiography system. Each examination was performed independently by a physician experienced in echocardiography, unaware of the results found by the alternative device. During the focused cardiac echocardiography, the following parameters were assessed: global cardiac systolic function, identification of ventricular enlargement or hypertrophy, assessment for pericardial effusion and estimation of the size and the respiratory changes of the inferior vena cava (IVC) diameter. RESULTS: One hundred fifty-one (151) patients were analyzed. With the tested PUD, the image quality was sufficient to perform focused cardiac ultrasonography in all patients. Examination using PUD adequately qualified with a very good agreement global left ventricular systolic dysfunction (κ = 0.87; 95%CI: 0.76-0.97), severe right ventricular dilation (κ = 0.87; 95%CI: 0.71-1.00), inferior vena cava dilation (κ = 0.90; 95%CI: 0.80-1.00), respiratory-induced variations in inferior vena cava size in spontaneous breathing (κ = 0.84; 95%CI: 0.71-0.98), pericardial effusion (κ = 0.75; 95%CI: 0.55-0.95) and compressive pericardial effusion (κ = 1.00; 95%CI: 1.00-1.00). CONCLUSIONS: In an emergency setting, this new ultraportable echoscope (PUD) was reliable for the real-time detection of focused cardiac abnormalities.

Acute myocardial infarction and multiple traumas: a dilemma of protocol.

Treatment of myocardial infarction is today governed by specific protocols. Angioplasty involves a therapeutical anticoagulation to prevent the risk of acute thrombosis. Acute myocardial infarction after a blunt trauma has been described, but there is no specific treatment recommendations extant, particularly weighing the risk of hemorrhage. In this report, we describe an adolescent boy who suffered from an acute myocardial infarction by dissection of the left anterior descending coronary after a car crash. He also presented with a subdural hemorrhage and a lung contusion, injuries, which both present a substantial risk of hemorrhage. After diagnosing the therapeutical problem, we describe our decisions regarding how we approached this case. We provide an algorithm of treatment coming from our experience of these cases with the hope it can help physicians in their future decisions.