Application of udder surface temperature by infrared thermography for diagnosis of subclinical mastitis in Holstein cows located in tropical highlands.

Several reports have indicated that udder surface temperature (UST) can be a useful indicator of subclinical mastitis (SCM). The objective was to evaluate UST by infrared thermography (IRT) as a diagnostic tool for SCM and intramammary infection (IMI), and to assess the influence of environmental conditions in the potential diagnosis of this disease in dairy cows located at high-altitude tropical regions. A total of 105 cows (397 quarters) from 3 dairy farms with mechanical and manual milking methods were enrolled in the study. Subclinical mastitis was diagnosed when quarter samples had a somatic cell count (SCC) ≥200 × 103 cells/mL, microbial growth (MG) was defined when a major pathogen (≥1 cfu/plate) or Corynebacterium spp. (≥10 cfu/plate) was isolated, and IMI was defined as the presence of MG and SCC ≥100 × 103 cells/mL. Infrared images were taken with a thermal camera placed 1 m away from the udder, and shots of the rear and left and right lateral view were made during the morning milking, before any manipulation of the udder and employing dark cardboard on the contralateral side to avoid artifacts in the background. A multilevel mixed effects linear regression model clustered within cows and herd was performed to evaluate the associations with UST. Clinical performance was evaluated using the Youden index to establish the optimum UST thresholds, which were set at 32.6°C for any case definition when milking was by hand, at 33.7°C for MG, and at 34°C for SCM and IMI in machine-milked quarters. Sensitivity (Se), specificity (Sp), area under curve (AUC), and positive likelihood ratio (+LR) were also assessed. Test agreement was assessed by kappa coefficient (κ). The UST of healthy quarters ranged between (95% CI) 32.4 and 32.6°C, lower than SCM quarters (n = 88) at 32.9°C (95% CI: 32.7-33.1 °C), MG quarters (n = 56) at 33.5°C (95% CI: 33.3-33.7°C), and IMI quarters (n = 50) at 33.5°C (95% CI: 33.2-33.7 °C). The UST was also related to the milking method: higher temperatures were observed for hand milking (n = 90) compared with machine milking (n = 185). No relation between environmental conditions such as wind speed, atmospheric temperature, relative humidity, and temperature-humidity index and UST were observed during this study. For hand milking, the optimal UST threshold was 32.6°C; for SCM, Se = 0.53, Sp = 0.89, AUC = 0.71, κ = 0.4; for MG, Se = 0.83, Sp = 0.93, AUC = 0.88, κ = 0.77; and for IMI, Se = 0.82, Sp = 0.92, AUC = 0.87, κ = 0.74. The machine milking threshold for SCM resulted in Se = 0.42, Sp = 0.97, AUC = 0.70, κ = 0.47; for MG, Se = 0.82, Sp = 0.89, AUC = 0.85, κ = 0.60; and for IMI, Se = 0.82, Sp = 0.98, AUC = 0.90, κ = 0.79. These findings suggest that UST determined by IRT is higher in machine-milked cows and in quarters with MG and IMI than in healthy quarters; therefore, UST by IRT is a reliable, clinically useful method for MG and IMI diagnosis.

Ciclo de vida y enemigos naturales de Mechanitis menapis (Lepidoptera: Ithomiini)

Las mariposas de la tribu Ithomiini son uno de los modelos biológicos más estudiados en años recientes en trabajos de biogeografía, taxonomía y evolución. Sin embargo, aun cuando la biología y distribución de sus especies es mejor conocida que la de otros grupos de mariposas, existen aspectos de la historia natural poco estudiados que permitirían un mejor entendimiento del comportamiento de sus poblaciones y las interacciones con su ambiente. En este trabajo, se estudió la historia natural de la mariposa, Mechanitis menapis mantineus Hewitson (Nymphalidae: Ithomiini) y sus enemigos naturales en el oeste de Ecuador. Para ello, se realizó la identificación de las plantas hospederas, se estudió el ciclo de vida y los factores de mortalidad de los estados inmaduros. Adicionalmente, se identificaron los parasitoides asociados a la especie en la zona de estudio. Se identificaron tres plantas hospederas, las cuales son nuevos registros para la especie en la región. Asimismo, se describió la morfología de los estados inmaduros y su tiempo de desarrollo. Finalmente, se identificaron los principales enemigos naturales y se describieron aspectos de la historia natural y comportamiento de Hyposoter sp. (Ichneumonidae: Campopleginae), principal parasitoide de larvas. Los resultados de este trabajo, proporcionan información esencial para la revisión taxonómica del género Mechanitis F. y para continuar el estudio de la interacción de esta especie con sus plantas hospederas y enemigos naturales.

Life cycle and natural enemies of Mechanitis menapis (Lepidoptera: Ithomiini). The butterflies of the Ithomiini tribe are one of the most-studied biological models of recent years in terms of biogeography, taxonomy, and evolution. However, even though their biology and distribution is better known than many other groups of butterflies, there are unknown aspects of their natural history that would improve our understanding of their behavior, population dynamics, and interactions with their environment. In this work, we studied the natural history of the butterfly Mechanitis menapis mantineus Hewitson (Nymphalidae: Ithomiini), and its natural enemies, in Western Ecuador. We identified three host plants, which are new records for the species in this region. We documented the life cycle and described the morphology of the immature stages, their development time, and studied the factors associated with mortality of these immature stages. Additionally, we identified the parasitoids associated with the species in the study area. In particular, we documented aspects of the natural history and behavior of Hyposoter sp. (Ichneumonidae: Campopleginae), the main parasitoid of the larvae. The results of this work provide essential information for the taxonomic revision of the genus Mechanitis F. as well as the continued study of the interactions between the butterfly, its host plants, and natural enemies.

Equine suspensory ligament and tendon explants cultured with platelet-rich gel supernatants release different anti-inflammatory and anabolic mediators.

The aim of this study was to evaluate the release of pro- and anti-inflammatory as well as anabolic mediators stimulated by a leukocyte-reduced platelet-rich gel supernatant (Lr-PRGS) and a leukocyte-reduced plasma supernatant (Lr-PL) at two concentrations (25 and 50%) on normal equine suspensory ligament explants (SLEs) and tendon explants (TEs). SLEs and TEs from six horses were independently incubated for 48 h with Lr-PRGS and Lr-PL at concentrations of 25 and 50%, respectively. Samples were collected from the incubated tissues at 1 h and 48 h, which were employed for ELISA determination of interleukin 1 beta (IL-1ß), tumor necrosis factor alpha (TNF-α), IL-4, IL-1 receptor antagonist (IL-1ra), platelet-derived growth factor isoform BB (PDGF-BB), transforming growth factor beta-1 (TGF-ß1, and hyaluronic acid (HA). Overall, 50% Lr-PRGS induced significantly less IL-1ß release than the other hemoderivatives in both tissues. At 48 h, both Lr-PRGS and 25% Lr-PL induced significantly higher TNF-α concentrations in SLEs when compared to TEs, whereas both Lr-PRGS concentrations induced significantly higher IL-4 concentrations in SLEs in comparison to TEs. IL-1ra release was not different between tissues. However, this cytokine was significantly higher in tissue explants cultured with both Lr-PRGS concentrations. HA concentration was lower in tissue explants cultured with all hemoderivatives at two concentrations when compared to the control group. The positive effects observed for ligaments and tendons treated with Lr-PRGS may be mediated by the inhibition of IL-1ß release of and increased release of IL-4 and IL-1ra. Furthermore, PDGF-BB could be a polypeptide responsible for mediating the release of anti-inflammatory cytokines in SLEs and TEs incubated with Lr-PRGS.

Contrasting patterns of Andean diversification among three diverse clades of Neotropical clearwing butterflies.

The Neotropical region is the most biodiverse on Earth, in a large part due to the highly diverse tropical Andean biota. The Andes are a potentially important driver of diversification within the mountains and for neighboring regions. We compared the role of the Andes in diversification among three subtribes of Ithomiini butterflies endemic to the Neotropics, Dircennina, Oleriina, and Godyridina. The diversification patterns of Godyridina have been studied previously. Here, we generate the first time-calibrated phylogeny for the largest ithomiine subtribe, Dircennina, and we reanalyze a published phylogeny of Oleriina to test different biogeographic scenarios involving the Andes within an identical framework. We found common diversification patterns across the three subtribes, as well as major differences. In Dircennina and Oleriina, our results reveal a congruent pattern of diversification related to the Andes with an Andean origin, which contrasts with the Amazonian origin and multiple Andean colonizations of Godyridina. In each of the three subtribes, a clade diversified in the Northern Andes at a faster rate. Diversification within Amazonia occurred in Oleriina and Godyridina, while virtually no speciation occurred in Dircennina in this region. Dircennina was therefore characterized by higher diversification rates within the Andes compared to non-Andean regions, while in Oleriina and Godyridina, we found no difference between these regions. Our results and discussion highlight the importance of comparative approaches in biogeographic studies.

Study of a Two-Step Centrifugation Protocol for Concentrating Cells and Growth Factors in Bovine Platelet-Rich Plasma.

There is a lack of information about the methods used for bovine platelet-rich plasma (PRP)/platelet-rich gel (PRG) procurement, including information on platelet (PLT), white blood cell (WBC) in PRP, and growth factor release from PRG supernatants. The aims of this study were to compare and to correlate the PLT, WBC, transforming growth factor beta-1 (TGF-ß1), and platelet-derived growth factor BB (PDGF-BB) concentrations in bovine whole blood, plasma, and four PRP layers and their respective PRG supernatants: A and B (obtained by a single centrifugation tube method at 720g/5 min) and C and D (obtained by a double centrifugation tube method, by using two centrifugation episodes at 720g/5 min). PLT and WBC counts were significantly higher in PRP-C, followed by whole blood, PRP-A, PRP-B, and PRP-D. TGF-ß1 concentrations were significantly higher in PRG-B supernatants and its correspondent PRP-B lysate when compared to the other PRG supernatants and plasma. Supernatants from PRG-A, PRG-B, and PRG-D had equivalent TGF-ß1 concentrations. PDGF-BB concentrations were not statistically different between the hemoderivatives. Significant Pearson correlations were noted between PLT counts and WBC counts (0.8) and between PLT counts and PLT distribution width (0.6). Further studies should be performed to assess the potential clinical applications of these PRPs.

North Andean origin and diversification of the largest ithomiine butterfly genus.

The Neotropics harbour the most diverse flora and fauna on Earth. The Andes are a major centre of diversification and source of diversity for adjacent areas in plants and vertebrates, but studies on insects remain scarce, even though they constitute the largest fraction of terrestrial biodiversity. Here, we combine molecular and morphological characters to generate a dated phylogeny of the butterfly genus Pteronymia (Nymphalidae: Danainae), which we use to infer spatial, elevational and temporal diversification patterns. We first propose six taxonomic changes that raise the generic species total to 53, making Pteronymia the most diverse genus of the tribe Ithomiini. Our biogeographic reconstruction shows that Pteronymia originated in the Northern Andes, where it diversified extensively. Some lineages colonized lowlands and adjacent montane areas, but diversification in those areas remained scarce. The recent colonization of lowland areas was reflected by an increase in the rate of evolution of species' elevational ranges towards present. By contrast, speciation rate decelerated with time, with no extinction. The geological history of the Andes and adjacent regions have likely contributed to Pteronymia diversification by providing compartmentalized habitats and an array of biotic and abiotic conditions, and by limiting dispersal between some areas while promoting interchange across others.

Influence of calcium salts and bovine thrombin on growth factor release from equine platelet-rich gel supernatants.

OBJECTIVE: To compare five activation methods in equine platelet-rich plasma (PRP) by determination of platelet-derived growth factor BB (PDGF-BB) and transforming growth factor beta 1 (TGF-ß1) concentrations in platelet-rich gel (PRG) supernatants. METHODS: Platelet-rich plasma from 20 horses was activated by calcium chloride (CC), calcium gluconate (CG), bovine thrombin (BT), and their combinations, BTCC and BTCG. Both growth factor concentrations in PRG supernatants were measured by ELISA and compared with plasma and platelet lysates (PL) over time. RESULTS: Growth factor concentrations were significantly lower in plasma and higher for all PRG supernatants. Platelet lysates contained a significantly lower concentration of PDGF-BB than PRG supernatants and a significantly higher concentration of TGF-ß1 than PRG supernatants. Clots from PRP activated with sodium salts were more stable over time and had significant growth factor release, whereas CC produced gross salt deposition. Significant correlations were noticed for platelet with leukocyte concentrations in PRP (rs: 0.76), platelet counts in PRP with TGF-ß1 concentrations in PRG supernatants (rs: 0.86), platelet counts in PRP with PDGF-BB concentrations in PRG supernatants (rs: 0.78), leukocyte counts in PRP with TGF-ß1 concentrations in PRG supernatants (rs: 0.76), and PDGF-BB concentrations with activating substances (rs: 0.72). CLINICAL SIGNIFICANCE: Calcium gluconate was the better substance to induce PRP activation. It induced growth factor release free from calcium precipitates in the clots. Use of BT alone or combined with calcium salts was not advantageous for growth factor release.

Into the Andes: multiple independent colonizations drive montane diversity in the Neotropical clearwing butterflies Godyridina.

Understanding why species richness peaks along the Andes is a fundamental question in the study of Neotropical biodiversity. Several biogeographic and diversification scenarios have been proposed in the literature, but there is confusion about the processes underlying each scenario, and assessing their relative contribution is not straightforward. Here, we propose to refine these scenarios into a framework which evaluates four evolutionary mechanisms: higher speciation rate in the Andes, lower extinction rates in the Andes, older colonization times and higher colonization rates of the Andes from adjacent areas. We apply this framework to a species-rich subtribe of Neotropical butterflies whose diversity peaks in the Andes, the Godyridina (Nymphalidae: Ithomiini). We generated a time-calibrated phylogeny of the Godyridina and fitted time-dependent diversification models. Using trait-dependent diversification models and ancestral state reconstruction methods we then compared different biogeographic scenarios. We found strong evidence that the rates of colonization into the Andes were higher than the other way round. Those colonizations and the subsequent local diversification at equal rates in the Andes and in non-Andean regions mechanically increased the species richness of Andean regions compared to that of non-Andean regions ('species-attractor' hypothesis). We also found support for increasing speciation rates associated with Andean lineages. Our work highlights the importance of the Andean slopes in repeatedly attracting non-Andean lineages, most likely as a result of the diversity of habitats and/or host plants. Applying this analytical framework to other clades will bring important insights into the evolutionary mechanisms underlying the most species-rich biodiversity hotspot on the planet.

Effects of sodium citrate and acid citrate dextrose solutions on cell counts and growth factor release from equine pure-platelet rich plasma and pure-platelet rich gel.

BACKGROUND: There is a lack information on the effects of the most commonly used anticoagulants for equine platelet rich plasmas (PRPs) elaboration on cell counts and growth factor release from platelet rich gels (PRGs). The aims of this study were 1) to compare the effects of the anticoagulants sodium citrate (SC), acid citrate dextrose solution A (ACD-A) and ACD-B on platelet (PLT), leukocyte (WBC) and on some parameters associated to platelet activation including mean platelet volume (MPV) and platelet distribution width (PDW) between whole blood, pure PRP (P-PRP) and platelet-poor plasma (PPP); 2) to compare transforming growth factor beta 1 (TGF-ß(1)) and platelet-derived growth factor isoform BB (PDGF-BB) concentrations in supernatants from pure PRG (P-PRG), platelet-poor gel (PPG), P-PRP lysate (positive control) and plasma (negative control); 3) to establish the possible correlations between all the studied cellular and molecular parameters. RESULTS: In all cases the three anticoagulants produced P-PRPs with significantly higher PLT counts compared with whole blood and PPP. The concentrations of WBCs were similar between P-PRP and whole blood, but significantly lower in PPP. The type of anticoagulant did not significantly affect the cell counts for each blood component. The anticoagulants also did not affect the MPV and PDW parameters. Independently of the anticoagulant used, all blood components presented significantly different concentrations of PDGF-BB and TGF-ß(1). The highest growth factor (GF) concentrations were observed from P-PRP lysates, followed by PRG supernatants, PPP lysates, PPG supernatants and plasma. Significant correlations were observed between PLT and WBC counts (ρ = 0.80), PLT count and TGF-ß(1) concentration (ρ = 0.85), PLT count and PDGF-BB concentration (ρ = 0.80) and PDGF-BB and TGF-ß(1) concentrations (ρ = 0.75). The type of anticoagulant was not correlated with any of the variables evaluated. CONCLUSIONS: The anticoagulants did not significantly influence cell counts or GF concentrations in equine PRP. However, ACD-B was apparently the worst anticoagulant evaluated. It is necessary to perform additional research to determine the effect of anticoagulants on the kinetics of GF elution from P-PRG.

Effects of the breed, sex and age on cellular content and growth factor release from equine pure-platelet rich plasma and pure-platelet rich gel.

BACKGROUND: There is no information on the effects of the breed, gender and age on the cellular content and growth factor (GF) release from equine pure-platelet rich plasma (P-PRP) and pure-platelet rich gel (P-PRG). The objectives of this study were: 1) to compare the cellular composition of P-PRP with whole blood and platelet poor plasma (PPP); 2) to compare the concentration of transforming GF beta 1 (TGF-ß1) and platelet derived GF isoform BB (PDGF-BB) between P-PRP treated with non-ionic detergent (P-PRP+NID), P-PRG (activated with calcium gluconate -CG-), PPP+NID, PPP gel (PPG), and plasma and; 3) to evaluate and to correlate the effect of the breed, gender and age on the cellular and GF concentration for each blood component. Forty adult horses, 20 Argentinean Creole Horses (ACH) and, 20 Colombian Creole Horses (CCH) were included. Data were analyzed by parametric (i.e.: t-test, one way ANOVA) and non parametric (Kruskal-Wallis test, Wilcoxon test) tests. Correlation analysis was also performed by using the Spearman and Pearson tests. A p ≤ 0.05 was set as significant for all tests. All the blood components were compared for platelet (PLT), leukocyte (WBC), TGF-ß1 and PDGF-BB concentrations. The effect of the breed, gender and age on these variables was analyzed. A P ≤ 0.05 was accepted as significant for all the tests. RESULTS: PLT counts were 1.8 and 0.6 times higher in P-PRP than in whole blood and PPP, respectively; WBC counts were 0.5 and 0.1 times lower in P-PRP, in comparison with whole blood and PPP, respectively. TGF-ß1 and PDGF-BB concentrations were 2.3 and 262 times higher, respectively, in P-PRG than in plasma, and 0.59 and 0.48 times higher, respectively, in P-PRG than in PPG. P-PRG derived from CCH females or young horses presented significantly (P < 0.001) higher PDGF-BB concentrations than P-PRG derived from ACH males or older horses. CONCLUSIONS: Our results indicated that P-PRP obtained by a manual method was affected by intrinsic factors such as the breed, gender and age. Equine practitioners should be aware that cellular and GF release from P-PRP/P-PRG could change according with the intrinsic variables associated with a patient in particular.

Evaluación de resultados de la osteotomía tipo Chevron con desplazamiento plantar para corrección de hallux valgus / Functional results of Chevron osteotomy with plantar displacement of the first metatarsal head for hallux valgus correction

Introducción: se describe una osteotomía tipo Chevron modificada para la corrección de hallux valgus adicionando el desplazamiento plantar de la osteotomía convencional. La motivación para llevar a cabo dicha modificación se basa en que la osteotomía de Chevron presenta grados variables de acortamiento del primer metatarsiano que pueden generar metatarsalgia por transferencia en pacientes con pie griego (index minus) en el cual el segundo metatarsiano es más largo. Materiales y métodos: se diseñó un estudio descriptivo prospectivo de los pacientes con diagnóstico de hallux valgus asociado a pie griego clínico o radiológico, metatarsalgia o callo plantar que fueron sometidos a cirugía de osteotomía tipo Chevron con desplazamiento plantar entre el 2008 y el 2010 en la Clínica del Campestre, Medellín. Todas las cirugías fueron realizadas por el mismo cirujano. En el posquirúrgico se evaluó la presencia y la intensidad del dolor, la persistencia del callo plantar y la presencia de arco transverso. Se usaron las escalas AOFAS y VASFA. Resultados: se evaluaron 27 cirugías, con una mediana de seguimiento de 5 meses (rango entre 1 y 18 meses). La mediana de edad de los pacientes fue 51 años (rango entre 31 y 68 años). La mayoría de los pacientes fueron de sexo femenino. En la evaluación prequirúrgica, 93% presentaban pie tipo griego clínico y 96% radiológico (index minus). El tipo de arco plantar transverso predominante fue el convexo en el 59% de los casos, cóncavo en el 22,5% y plano en el 18,5%. El 78% presentaban metatarsalgia; 85%, callo plantar, y 29%, deformidad del segundo dedo. La mediana de los ángulos IM y MTF fue 11° (rango entre 6° y 16°) y 27,5° (rango entre 13° y 38°), respectivamente. En la evaluación posquirúrgica, 7 casos continuaban presentando metatarsalgia ocasional y 1 callo plantar. El 95% de los casos recuperaron o mantuvieron el arco transverso; el 86% no tenían limitación o ésta era mínima con el uso del calzado. En cuanto a la actividad, 80% no presentaban limitaciones de ningún tipo. Discusión: los resultados obtenidos en esta serie de pacientes muestran hallazgos positivos a corto plazo por la pérdida del callo plantar y la recuperación del arco transverso con mejoría de la sintomatología.

Abscesos pleurales producidos por Staphylococcus aureus y Streptococcus spp. beta-hemolítico como secuela de pleuroneumonía en un caballo.Reporte de un caso / Abscessos pleurais causados por Staphylococcus aureus e Streptococcus spp. beta-hemolítico no rescaldo de pleuropneumonia em um cavalo. Relato de um caso / Pleural abscesses produced by Staphylococcus aureus and beta-haemolytic Streptococcus spp. as a sequel of pleuropneumonia in a horse. A case report

Los abscesos pleurales son una complicación frecuente en caballos con pleuroneumonía. Su tratamiento resulta difícil, requiere tiempo y dinero. Un caballo criollo argentino, castrado, de 13 años de edad presentó una pleuroneumonía grave que posteriormente se complicó con abscesos pleurales. El uso de anti-inflamatorios no esteroidales (fenilbutazona), una antibioterapia agresiva (ceftiofur, ampicilina y gentamicina), el drenaje eco-guiado y el lavado de los abscesos con una solución de yodopovidona al 0.05% permitió la recuperación exitosa del paciente de este reporte.

Pleural abscesses are a frequent complication in horses with pleuropneumonia. Their treatment is not easy, and requires time and it is expensive. A 13 years old creole argentine gelding presented a severe pleuropneumonia subsequently complicated with pleural abscess. The use of nonsteroidal antiinflammatory drugs (fenylbutazone), an aggressive antibiotherapy (ceftiofur, ampicillin, and gentamicin), echoguide drainage and lavage with yodopovidone to 0.05% of the abscesses permitted the successful recuperation of the patient of this case report.

Os abscessos pleurais são uma complicação comum em cavalos com pleuropneumonia. Seu tratamento é difícil e requer tempo e dinheiro. Um cavalo crioulo argentino, castrado, 13 anos de idade apresentou uma grave pleuropneumonia que posteriormente se tornou complicado com abscesso pleural. O uso de não-esteróides antiinflamatórios (fenilbutazona), uma agressiva terapia antibiótica (cetfiofur, ampicilina e gentamicina), eu drenagem eco-guiado e da lavagem dos abscessos com uma solução de yodopovidona de 0.05%, permitiu o sucesso da recuperação do paciente deste relato de caso.