Temporal Release and Denature of Several Mediators in Pure Platelet-Rich Plasma and Temperature-Induced Platelet Lysates Derived from a Similar Bovine Platelet Concentrate.

There is scarce information about bovine platelet-rich plasma/platelet-rich gel (PRP/PRG) and related hemocomponents (HCs), such as platelet lysates (PLs), including growth factor (GF) and cytokine concentrations, and how the stability of these biomolecules could be affected by time and temperature. This study aimed to evaluate the release and stability of transforming growth factor beta 1 (TGF-ß 1), interleukin 4 (IL-4), and tumor necrosis factor alpha (TNF-α) contained in bovine pure PRP (P-PRP) and temperature-induced PL (TIPL) coming from a similar platelet concentrate (PC) at 4 and 37°C at 3 and 96 h. Platelet concentrates (PCs) presented a 1.7-fold concentration of platelets (PLTs) with negligible counts of white blood cells (WBCs) when compared to the counts for these cells in whole blood. TGF-ß 1 concentrations were significantly lowest in plasma followed by TIPL, chemical-induced PL (CIPL), and P-PRP. IL-4 and TNF-α concentrations did not differ between HCs. TGF-ß 1 concentrations were negatively affected in P-PRPs stored at 4°C at 3 and 96 h, whereas those from P-PRP maintained at 37°C presented similar concentrations to TIPL stored at both temperatures over time. IL-4 and TNF-α concentrations were not affected by time or temperature in any of the HCs evaluated. Pure PRGs released additional quantities of GF and cytokines over time when compared with HCs stored over 96 h at 4 and 37°C. The method, either chemical or physical, used for platelet activation or damage produces a different GF and cytokine release pattern, which makes to each evaluated HCs different despite they come from a similar bovine PC. P-PRP activated with calcium gluconate and maintained at 37°C, which polymerizes in P-PRG, showed the best GF and cytokine release/denature profile compared with the rest of the HCs evaluated.

Intramammary treatment using allogeneic pure platelet-rich plasma in cows with subclinical mastitis caused by Gram-positive bacteria.

The aims of the study were (1) to compare the cure risk of intramammary treatment of pure platelet rich plasma (P-PRP) or cefquinome sulfate (CS) in cows with subclinical mastitis (SCM) caused by Gram-positive bacteria, evaluated via somatic cell count (SCC) and the microbiological analysis of milk; (2) to compare the inflammatory/anti-inflammatory response of mammary gland to both treatments through the analyses of interleukins (IL), interferon gamma (IFN-γ), and tumour necrosis factor alpha (TNF-α) in milk. A non-inferiority randomized clinical trial was conducted. The null hypothesis was that cure risk in the experimental group (P-PRP) was inferior to the reference group (CS). A total of 103 cows were selected according to SCC and presence of Gram-positive bacteria, 49 cows were treated with CS and 54 cows were treated with P-PRP after determination of its cellular and molecular quality control. Cure was assessed by milk analyses at day 21 and 22 after treatment. Cows that remained with SCM were retreated at day 26, and cure assessed at day 47 and 48. Overall, bacteriological cure was observed in 16 cows (30%) of the P-PRP group, and 35 cows (71%) in CS group. Staphylococcus aureus cure risk was higher in CS group, but inconclusive for Streptococcus spp. The mean SCC increased in relation to time only in the P-PRP group. A direct relation between time and treatment for IL-1, IL-2, and IL-6 was observed, while no differences were observed for IL-4. Furthermore, IL-1 and IL-2 increased in cows treated twice in both groups. IL-8, IFN-γ, and TNF-α showed a significant interaction between time and treatment. IFN-γ concentration was lower in the P-PRP group compared to the CS on days 0 and 22. Leukocyte counts were lower in P-PRP when compared to whole blood. TGF-ß1 and PF4 concentrations were higher in platelet lysates in comparison to P-PRGS and plasma. Moreover, PDGF-BB concentration was significantly higher in platelet lysates in comparison to plasma. Results obtained in this study demonstrate that SCM treated with PRP showed a lower rate of bacteriologic cure when compared to animals treated with CS.

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.

Características sociodemográficas y clínicas de pacientes atendidos en urgencias con diagnóstico de cáncer de novo / Socio-demographic and clinical characteristics of patients with de novo cancer diagnosis, admitted trough emergency services

Objetivo: caracterizar sociodemográfica y clínicamente los pacientes que después de ingresar a servicios de urgencias de dos centros de tercer nivel de atención de la ciudad de Manizales (Colombia), se les diagnosticó alguno de los tipos de cáncer más frecuentes según el Registro Poblacional de Cáncer de Manizales. Materiales y métodos: con base a la información aportada por el Registro Poblacional de Cáncer de Manizales entre los años 2008 y 2012, se escogieron los cinco tipos de cáncer más frecuentes en cada género para la realización del estudio. Fueron incluidos 116 casos de los que ingresaron por servicios de urgencias para extraer información a partir de las historias clínicas, la cual posteriormente fue tabulada y analizada mediante estadística descriptiva. Resultados: la mayoría los pacientes provenían de Manizales (60,3%), y pertenecían al régimen contributivo de seguridad social (50,0%). El promedio de edad fue de 72,4 años. El promedio de duración de los síntomas antes de consultar fue de 2,3 meses y los cánceres más frecuentes fueron el gástrico (41,4%), de colon (25,9%), CP (23,3%), cérvix (6%) y recto (3,5%) en estadios avanzados. Conclusiones: se continúan diagnosticando casos de cáncer de novo en estadio tardío, al ingresar por los servicios de urgencias. Resulta muy importante conocer la epidemiología de estos tipos de cáncer en cada una de las regiones y mejorar la formación médica con el fin de implementar estrategias para disminuir su aparición..(AU)

Objective: to characterize sociodemographicly and clinically those patients, who after admission by the emergency services of two tertiary level centers in Manizales (Colombia), were diagnosed with some of the most frequent cancers according to the Manizales Population Cancer Registry. Materials and methods: based on the information provided by the Manizales Population Cancer Registry between the years of 2008 to 2012, the five most frequent types of cancer in each gender were chosen for the study. There were 116 cases of those admitted through the emergency services for data extraction from the clinical record, which was later tabulated and analyzed using descriptive statistics. Results: most of the patients came from Manizales (60.3%), covered by the contributory social security scheme (50.0%), with a mean age of 72.4 years. The mean duration of symptoms before consulting the emergency service was 2.3 months. The most frequent cancers were gastric (41.4%), colon (25.9%) lung (23.3%), cervix (6%) and rectal cancer (3.5%) in advanced stages. Conclusions: new cases of late-stage cancer are still being diagnosed upon admission to emergency services. It is important to know the epidemiology of these cancers in each of the regions and to improve medical training in order to implement strategies to reduce their presentation..(AU)

Blood ß-hydroxybutyrate concentrations and early lactation management strategies on pasture-based dairy farms in Colombia.

The increasing global demand for food requires sustainable solutions to close the gap in agricultural yield between industrialized and non-industrialized countries. Our objectives in this cross-sectional study were to: 1) characterize farm populations, milk yield, and early lactation management strategies of dairy cows in three different regions of Colombia, and 2) determine the association of these management strategies with blood ß-hydroxybutyrate (BHB) concentrations in the first 42 days in milk (DIM). Dairy herds (n = 56) in the Antioquia, Caldas, and Cundinamarca regions of Colombia were visited once from May through July 2018. A survey was administered to farm owners to collect demographic, management, and herd nutrition information. Blood samples from dairy cows (n = 880) between calving and 42 DIM were used to measure blood BHB concentration. Associations between management and nutritional strategies and blood BHB concentration were examined using mixed models. Prevalence of hyperketonemia was calculated as the number of samples with BHB concentration ≥1.2 mmol/L divided by the total number of samples. The estimated diet composition for early lactation dairy cows was 65.5% pasture and 31.8% commercial concentrates. The farm median milk yield, protein concentration, and fat concentration were 21.0 kg (range = 13.1-36 kg), 3.2% (range = 2.7-4.1%), and 3.5% (range = 3.0-4.1%), respectively. Milk yield least squares means (95% confidence interval; CI) differed by region: 21.7 (20.3, 23.2), 18.5 (17.0, 20.2), and 20.3 (18.5, 22.4) kg in Antioquia, Caldas, and Cundinamarca, respectively. Median blood BHB concentration was 0.5 and ranged from 0.1-4.4 mmol/L; blood BHB concentration was not different among the three regions. Pasture fertilization, increased parity, and BCS were associated with changes in blood BHB concentration. The overall prevalence of hyperketonemia was 4.5%. Geographical region affected the prevalence of hyperketonemia at 2.5%, 4.0%, and 10.2% in Antioquia, Caldas, and Cundinamarca, respectively. Mean stocking density (95% CI) was greater in Cundinamarca than Antioquia or Caldas at 3.3 (2.2, 5.0), 2.8 (2.1, 3.9) and 1.7 (1.2, 2.6) animals per ha, respectively, and was associated with hyperketonemia prevalence. Farms that abruptly stop milking cows at dry-off had 80% of the hyperketonemia events in the study. Pasture-based dairies in Colombia had lower blood BHB concentrations and estimated milk yield compared with confined production systems in temperate zones. However, geographical region, stocking density, and abrupt cessation of milking at dry-off were associated with prevalence of hyperketonemia in pasture-based dairies.

Potential group B Streptococcus interspecies transmission between cattle and people in Colombian dairy farms.

Group B Streptococcus (GBS), is a leading cause of neonatal death and an emerging pathogen in adults. Additionally, GBS is a bovine pathogen causing intramammary infections. The likelihood of GBS interspecies transmission is largely unknown. We explored the potential transmission of GBS between cattle and people on dairy farms in Colombia and compared the antimicrobial resistance (AMR) profiles of isolates from both host species. Across 33 farms, throat swabs and rectal swabs were collected from 191 people, and rectal swabs and composite milk samples from 2092 cattle, yielding 60 human isolates and 301 bovine isolates. The majority (64%) of isolates belonged to shared sequence types (ST). Sequence type (ST) 1 was the most common strain in both host species, suggesting that interspecies transmission may be possible. Two members of the bovine-specific clonal complex 61/67 were detected in human samples (ST718 and ST1175), providing evidence for the lack of genuine species barriers. Apparent prevalence of penicillin resistance was surprisingly high in human and bovine isolates. Further investigation of this phenomenon is needed and could lead to modification of standard testing and treatment recommendations in human and veterinary medicine.

Streptococcus agalactiae is not always an obligate intramammary pathogen: Molecular epidemiology of GBS from milk, feces and environment in Colombian dairy herds.

For many years Streptococcus agalactiae has been considered an obligate intramammary and strictly contagious pathogen in dairy cattle. However, recent reports of S. agalactiae isolation from extramammary sources have contradicted that premise. To gain further insight into the epidemiology of S. agalactiae infection in cattle, we examined its distribution and heterogeneity of strains in bovine milk, bovine feces, and the environment in Colombian dairy farms. First, a longitudinal study was conducted at herd level in 152 dairy herds. Bulk tank milk samples from each herd where collected twice a month for six months. A follow-up study with a cross sectional design at the cow level was conducted in a subset of 25 farms positive for S. agalactiae. Cow-level milk samples from 1712 lactatting cows and 1545 rectal samples were collected, as well as 120 environmental samples. Samples were used for S. agalactiae detection and genotyping using Multi Locus Sequence Typing. Results showed sporadic rather than repeated isolation of S. agalactiae from bulk tank milk in 40% of the positive herds, challenging the idea that S. agalactiae is a highly contagious pathogen causing chronic infections. S. agalactiae was isolated from rectal or environmental samples in 32% and 12% of cross-sectional study farms, respectively, demonstrating that the bacteria can survive in extramammary sources and that S. agalactiae is not an obligate intramammary pathogen. The same strain was isolated from rectal and bulk tank milk samples in eight farms, suggesting that fecal shedding is frequent, and contributes to the presence of S. agalactiae in bulk tank. High within-herd heterogeneity of strains was found, which is distinct from the situation in developed dairy industries. These new epidemiological findings should be considered to adjust surveillance and control recommendations for S. agalactiae.

Staphylococcus aureus Isolates from Bovine Mastitis in Eight Countries: Genotypes, Detection of Genes Encoding Different Toxins and Other Virulence Genes.

Staphylococcus aureus is recognized worldwide as one of the major agents of dairy cow intra-mammary infections. This microorganism can express a wide spectrum of pathogenic factors used to attach, colonize, invade and infect the host. The present study evaluated 120 isolates from eight different countries that were genotyped by RS-PCR and investigated for 26 different virulence factors to increase the knowledge on the circulating genetic lineages among the cow population with mastitis. New genotypes were observed for South African strains while for all the other countries new variants of existing genotypes were detected. For each country, a specific genotypic pattern was found. Among the virulence factors, fmtB, cna, clfA and leucocidins genes were the most frequent. The sea and sei genes were present in seven out of eight countries; seh showed high frequency in South American countries (Brazil, Colombia, Argentina), while sel was harboured especially in one Mediterranean country (Tunisia). The etb, seb and see genes were not detected in any of the isolates, while only two isolates were MRSA (Germany and Italy) confirming the low diffusion of methicillin resistance microorganism among bovine mastitis isolates. This work demonstrated the wide variety of S. aureus genotypes found in dairy cattle worldwide. This condition suggests that considering the region of interest might help to formulate strategies for reducing the infection spreading.

Errores conceptuales de estadística más comunes en publicaciones científicas / Common conceptual statistical mistakes in scientific literature / Equívocos comuns de estatísticas em publicações científicas

Resumen Este trabajo proporciona información actualizada sobre los errores estadísticos que comúnmente se cometen en la literatura científica y como estos podrían ser evitados. El objetivo principal es una revisión de manera conceptual de los errores estadísticos frecuentemente observados, así como defectos y trampas en la ciencia médica y veterinaria para ayudar a los investigadores a producir resultados estadísticamente correctos en sus futuras investigaciones. Al mismo tiempo, esta revisión podría ayudar a que los lectores de revistas científicas identifiquen análisis estadísticos o presentación de datos cuestionables y puedan estimar lo qué los autores habrían concluido si hubieran utilizado métodos estadísticos apropiados.

Abstract This work seeks to provide up-to-date information on commonly made statistical mistakes and statistical reports in scientific papers, and how these can be avoided. The main goal is to comprehensively review frequently observed statistical errors, flaws and pitfalls in medical and veterinary science in order to help researchers to produce statistically correct output in their future reports. At the same time, it can help readers to identify questionable statistical analysis, and estimate what the authors would have concluded when appropriate statistical methods have been used.

Resumo Este trabalho procura fornecer uma atualização sobre os erros estatísticos comumente cometidos na literatura científica e como eles podem ser evitados. O principal objetivo é analisar de uma maneira conceitual defeitos observados frequentemente, erros estatísticos e armadilhas na ciência médica e veterinária para ajudar os pesquisadores a produzir resultados estatisticamente corretos em futuras investigações. Ao mesmo tempo, esta revisão pode ajudar os leitores de revistas científicas a identificar análise estatística ou apresentação de dados questionáveis e estimar o que os autores teria concluído teve eles usaram métodos estatísticos adequados.

Bulk tank milk selenium and its association with milk production parameters in Canadian dairy herds.

An observational study was conducted in 63 Canadian dairy farms to evaluate the association of bulk tank milk selenium (Se) concentration (BTSe) with average daily milk yield, milk fat and milk protein, bulk tank somatic cell count (BTSCC), and the probability of being a Staphylococcus aureus-positive herd. Bulk tank milk samples collected between March 2007 and February 2008 were evaluated for BTSCC, S. aureus culture status, and BTSe. Mean BTSe was 0.51 ± 0.15 µmol/L; no herds were classified as deficient or marginal based on BTSe. Bulk tank milk Se was unconditionally associated with milk production; however, adjusting by region indicated a confounding effect of this variable. There was no relationship between BTSe and BTSCC. Higher values of BTSe were associated with lower risk of being a S. aureus-positive herd, possibly as a result of a more robust udder immune response, or as a result of providing Se at a higher rate as one component of an udder health program that reduces risk of being S. aureus positive.