Effect of the addition of antifreeze protein type I on the quality of post-thawed domestic cat epididymal sperm.

Cryopreservation of domestic cat semen is mainly performed as a model for the establishment of endangered wild feline protocols. The supplementation of antifreeze protein type I (AFP I) to cryopreservation medium has shown improvement in frozen-thawed sperm quality in other species, but its effect on cat semen has not yet been tested. This study aimed to assess the addition of AFP I to cryopreservation medium in domestic cats. Sperm was obtained from the cauda epididymis of orchiectomized cats; sperm was then pooled in Tris buffer and allocated into three treatments, according to AFP I final concentration: 0 (control), 0.1, and 0.5 µg/ml. Nine replicates were cryopreserved in a two-step protocol and subsequently thawed at 37°C for 30 s. There was no difference (P > 0.05) among the control, 0.1 and 0.5 µg/ml groups for parameters such as motility, vitality, functional membrane integrity, mature chromatin, normal morphology, and sperm binding to egg perivitelline membrane. In the 0.5 µg/ml group only, percentages of live sperm with intact acrosome and of sperm with most inactive mitochondria (DAB III) showed a significant reduction, along with a tendency (P = 0.053) to an increase in the percentage of sperm with most active mitochondria (DAB II). In conclusion, the supplementation of 0.1 and 0.5 µg/ml of AFP I did not promote consistent beneficial effects on the overall sperm cryotolerance in domestic cats.

A case of a diabetic woman with an uncontrollable arm.

We present the case of an 83-year-old female patient with left-side hemichorea and hemiballism associated with poorly controlled type 2 diabetes mellitus. Hyperintensity in the right basal ganglia was evident in T1-weighted brain magnetic resonance. Clinical and radiological findings were consistent with diabetic striatopathy.

Growth dynamic of Leptospira spp. from Sejroe serogroup in different media formulae.

The culturing of Leptospira strains from bovine clinical samples is challenging and has resulted in some gaps in securing an epidemiological understanding. Strains related to chronic reproductive leptospirosis in cattle belong to the Sejroe serogroup - not only Hardjoprajitno and Hardjobovis but also Guaricura genotypes. This study analyses the growth of Leptospira strains from serogroup Sejroe in different culture media, with the aim of suggesting better culturing approaches. To meet this objective, two culture media were applied: EMJH and T80/40/LH. In addition, three different cocktails of selective agents were chosen. The combinations of medium and selective additives resulted in 10 different tested formulae. The poor performance of Hardjobovis in EMJH indicated that its growth may represent a possible bias when culturing these strains from bovine samples. The most efficient medium for culturing Hardjobovis was T80/40/LH, while T80/40/LH medium + STAFF combination proved to be the best choice for growth, being recommended for obtaining a higher number of these strains from bovines.

Association between teat-end hyperkeratosis and mastitis in dairy cows: A systematic review.

A systematic review was conducted to elucidate the role of teat-end hyperkeratosis (THK) as a risk factor for clinical mastitis (CM) or subclinical mastitis (SCM). Scientific papers on the subject were identified by means of a database search. All types of peer-reviewed analytical studies, observational or experimental and published in English, could be included in the review, regardless of publication year. Of 152 identified records, 18 articles were selected, of which 8 were prospective cohort studies, 9 were cross-sectional, and 1 was a hybrid case-control study. Internal validity of studies was assessed using a score system ranging from 0 to 6, based on design, risk of bias, and statistical methods. The most frequent study limitation was improper use of statistical methods to avoid confounding of associations between THK and CM or SCM. The 3 studies that used CM as outcome (all with high validity scores) showed positive associations with THK (especially severe), although the magnitude and statistical significance of the estimates differed among them. Most studies that used SCM as the primary outcome (based on microbiological examination of milk) reported that only severe THK was associated with SCM. Two studies with high validity scores reported moderate to strong associations between severe THK and incidence or prevalence of Staphylococcus aureus intramammary infection. Two studies with high validity scores reported that only severe THK was associated with the risk of somatic cell count (SCC) ≥200,000 cells/mL and increased mean SCC, respectively. Although 4 cross-sectional studies reported positive associations between THK and SCC, these associations were possibly spurious because confounding factors, such as parity, were not considered in the analyses. Results of the reviewed studies suggest that severe THK is a risk factor for both CM and SCM, as defined by microbiological examination of milk, SCC thresholds, or mean SCC. The effect of severe THK on both contagious (especially Staphylococcus aureus) and environmental CM or SCM emphasizes the importance of teat health for mastitis control. Four studies demonstrated that quarters with mild THK had lower prevalence of intramammary infection or lower mean SCC than quarters with no THK, suggesting that development of mild THK, as a physiological response to milking, can have a protective effect. Dairy consultants should focus on monitoring and minimizing occurrence of severe THK to prevent CM and SCM.

HHRISK: A code for assessment of human health risk due to environmental chemical pollution.

Chemical environmental pollution is currently one of the most concerning environmental problem on a global scale, due to the high risks posed to ecological systems and human health. Risk assessment methodologies are valuable tools for preventive management and the mitigation of human health risks. However, the application of these methodological tools involves several steps and the knowledge of many variables, which can hinder its correct implementation. The main objective of this work was the development of the computational code for human health risk assessment: HHRISK (Human Health Risk). This code allows for an agile and accurate risk assessment based on the methodology established by the U.S. Environmental Protection Agency (U.S. EPA). Different from other published methods, the HHRISK code includes a new spatiotemporal matrix for the analysis of the aggregated risk (for multiple exposure pathways) and the cumulative (for exposure to multiple chemicals). HHRISK was applied to two case studies published dealing with the assessment of risk to human health through exposure to toxic metals, obtaining satisfactory results. The concordance between the average results obtained with the HHRISK and those reported by the authors confirm the validity of the implemented model. The inclusion of a greater spatiotemporal detail of the risks allowed to carry out a more accurate analysis and to propose new subsidies for a more efficient risk mitigation management by affected place and period of time.

Reduced susceptibility in leptospiral strains of bovine origin might impair antibiotic therapy.

Leptospirosis is a worldwide zoonotic disease determined by pathogenic spirochetes of the genus Leptospira. The control of bovine leptospirosis involves several measures including antibiotic treatment of carriers. Despite its importance, few studies regarding antimicrobial susceptibility of strains from bovine origin have been conducted. The aim of this study was to determine the in vitro susceptibility of Leptospira strains obtained from cattle in Rio de Janeiro, Brazil, against the main antibiotics used in bovine veterinary practice. A total of 23 Leptospira spp. strains were investigated for minimum inhibitory concentrations (MICs) and minimum bactericidal concentrations (MBCs) using broth macrodilution. At the species level, there were not differences in MIC susceptibility except for tetracycline (P < 0.05). Nevertheless, at the serogroup level, differences in MIC were observed among Sejroe strains, mainly for ceftiofur, doxycycline and in MBC for streptomycin (P < 0.05). One strain presented MBC values above maximum plasmatic concentration described for streptomycin and was classified as presenting reduced susceptibility. Efficacy of antimicrobial therapy on bovine leptospirosis could be compromised due to occurrence of infection by Leptospira strains presenting reduced susceptibility.

Diagnostic accuracy of Somaticell, California Mastitis Test, and microbiological examination of composite milk to detect Streptococcus agalactiae intramammary infections.

The objectives of this study were to estimate the accuracy of Somaticell (Idexx Laboratories Inc., Westbrook, ME), California Mastitis Test (CMT), and microbiological examination of composite milk (MEC) to diagnose Streptococcus agalactiae intramammary infections (IMI), and to assess the agreement between Somaticell and CMT to detect these infections. A secondary objective was to estimate quarter- and cow-level prevalence of S. agalactiae IMI in the herds included in the study. Seven farms were included in the study. The CMT was performed and aseptic milk samples were collected from all quarters of all lactating cows. Composite milk samples were produced in the laboratory by mixing milk from all quarters of each sampled cow. The Somaticell test was performed on a subset of S. agalactiae-positive (n = 167) and S. agalactiae-negative (n = 152) quarter milk samples. Microbiological examination of quarter milk samples (MEQ) was considered the reference test for diagnosing S. agalactiae IMI. The accuracy of all tests at various thresholds was estimated using Bayesian latent class models. Apparent prevalence of S. agalactiae IMI was 15.8% (n = 184/1,164) at the quarter level (based on MEQ) and 28.5% (n = 83/291) at the cow level (based on MEC). True prevalence, as determined by Bayesian models, was 13.0% [95% credible interval (CR): 6.4-24.4%] at the quarter level, and 25.6% (95% CR: 15.3-39.5%) at the cow level. At the cow level (n = 285), sensitivity and specificity of MEC were 95.6 and 99.5%, respectively. The accuracy of Somaticell (n = 319 quarters) to identify S. agalactiae-infected quarters was 75.4, 86.4, 88.9, 89.4, and 91.0% at thresholds of 98,000, 147,000, 205,000, 244,000, and 282,000 cells/mL, respectively. The accuracy of CMT was 87.6, 90.7, 90.8, and 87.4% at thresholds of trace, 1, 2, and 3, respectively. The areas under the receiver operating characteristic curve for Somaticell and CMT were 94.5% (95% confidence interval: 91.8-97.2%) and 92.0% (88.6-95.4%), respectively. At the tested thresholds, the sensitivity of Somaticell ranged from 94.9 to 99.5% to detect S. agalactiae IMI, and specificity ranged from 48.1 to 87.1%. The sensitivity of Somaticell at the lowest threshold (69,000 cells/mL; sensitivity = 99.9%; 95% CR: 98.2-100%) was higher than that of CMT at any tested threshold. Results of this study could be used at the farm level to reduce the use of antimicrobials and reach specific goals in S. agalactiae eradication programs.

Bioaccumulation of nickel in tomato plants: risks to human health and agro-environmental impacts.

Anthropogenic activities such as agriculture, industry, and mining have contributed significantly to the accumulation of heavy metals in the soil, which in turn cause problems to human health and to the environment. The present work aims to study the effects of nickel (Ni) on the development of tomato plants, the risks to human health associated to the consumption of contaminated tomatoes, and the consequences to the environment. The experiment was carried out in greenhouse environment for a period of 120 days, and the plants were cultivated in soils with four different concentrations of Ni: 0, 35, 70, and 105 mg kg-1. The concentration of nickel in each part (root, stem, leaf, and fruit) of the tomato plant was measured at four different stages of the cycle: 30, 60, 90, and 120 days, by inductively coupled plasma optical emission spectrometer (ICP-OES). At the end of the cycle, the concentration of certain macro- and micronutrients was also determined and related to the corresponding Ni concentration in the soil. The distribution of Ni in the parts of the plant was analyzed from the bioaccumulation factor temporal behavior. Nickel concentrations found in the fruit were too low to pose a risk to human health. As a result of this research, it was verified that soils with nickel concentrations close to 70 mg kg-1, which is the limit established by the CONAMA resolution (420/2009), may actually represent an optimum concentration value for the development of tomato plants. It also increases productivity per plant and reduces the use of resources such as water and agricultural inputs.

Predictive value of ophthalmic artery Doppler velocimetry in relation to development of pre-eclampsia.

OBJECTIVE: To test the hypothesis that ophthalmic artery Doppler velocimetry is predictive of the development of pre-eclampsia (PE). METHODS: This was a prospective cohort study that included pregnant women in the second trimester who had risk factors for PE. Seven ophthalmic artery Doppler parameters, in addition to uterine artery (UtA) Doppler and clinical variables, were investigated for their prognostic value with respect to PE. RESULTS: A total of 347 women were recruited, of whom 40 developed PE. A comparison of the mean ophthalmic artery Doppler parameter values between women with and those without PE showed statistically significant differences in several parameters: peak systolic velocity, end-diastolic velocity, mean velocity, peak mesodiastolic velocity (PMDV) and peak ratio. After adjusting for confounding variables, only PMDV remained statistically significant (P < 0.001), with an area under the receiver-operating characteristics curve (AUC) of 0.73. The best cut-off for predicting PE was a PMDV of > 22.11 cm/s, with sensitivity of 70%, specificity of 75%, positive likelihood ratio of 2.8, negative likelihood ratio of 0.4, positive predictive value of 28% and negative predictive value of 95%. The AUC increased from 0.72 to 0.78 when the PMDV was incorporated into a prediction model based on clinical variables, demonstrating that this marker increased the discriminatory capability of the model. The performance of ophthalmic artery Doppler was similar to that of UtA Doppler for predicting PE. Additionally, the AUC increased significantly from 0.82 to 0.88 when the PMDV was incorporated into the model containing clinical variables and UtA Doppler indices. CONCLUSION: A high ophthalmic artery PMDV in the second trimester of pregnancy is an independent predictor of PE that increases the discriminatory ability of clinical markers, as well as of models that include clinical variables and UtA Doppler indices.

The effect of urbanization on species composition and trophic guilds of bats (Mammalia, Chiroptera) in the Brazilian Savanna.

Urban environments present less environmental heterogeneity in relation to the natural ones, affecting the biodiversity of bats and the ecological processes in which they participate. In this way, we will identify how urbanization influences the structure of bat communities in the municipality of Goiânia, Goiás, Brazil. We compared species composition, guilds and bat richness in a gradient that crossed urban, semi-urban and natural areas in the municipality of Goiânia, contained in the Cerrado biome. We captured a total of 775 bats of 16 species distributed in three families. Urban areas had a higher species abundance, while semi-urban areas had a higher species richness. The three types of environments have different compositions, the urban one being more homogeneous, the fauna in these areas is composed of generalist species, which benefit from this process. The diversity present in semi-urban areas is a consequence of the intersection between urban and natural fauna, which is why urban expansion needs to occur in a planned manner to minimize the impacts of this process and ensure the maintenance of biodiversity.

Tributyltin (TBT) toxicity: Effects on enteric neuronal plasticity and intestinal barrier of rats' duodenum.

Tributyltin (TBT) is a biocide used in the formulation of antifouling paints and it is highly harmful. Despite the ban, the compound persists in the environment, contaminating marine foodstuffs and household products. Therefore, considering the route of exposure to the contaminant, the gastrointestinal tract (GIT) acts as an important barrier against harmful substances and is a potential biomarker for understanding the consequences of these agents. This work aimed to evaluate histological and neuronal alterations in the duodenum of male Wistar rats that received 20 ng/g TBT and 600 ng/g via gavage for 30 consecutive days. After the experimental period, the animals were euthanized, and the duodenum was intended for neuronal histochemistry (total and metabolically active populations) and histological routine (morphometry and histopathology). The results showed more severe changes in neuronal density and intestinal morphometry in rats exposed to 20 ng/g, such as total neuronal density decrease and reduction of intestinal layers. In rats exposed to 600 ng/g of TBT, it was possible to observe only an increase in intraepithelial lymphocytes. We conclude that TBT can be more harmful to intestinal homeostasis when consumed in lower concentrations.

Far-UV-C irradiation promotes synergistic bactericidal action against adhered cells of Escherichia coli and Staphylococcus epidermidis.

The contamination of indoor areas is a global health problem that can cause the dispersion of infectious diseases. In that sense, it is urgent to find new strategies applying a lower concentration of the traditional chemicals used for cleaning and disinfection. Ultraviolet radiation (UV), in particular far-UV-C (200-225 nm), has emerged as a successful, powerful, easy-to-apply, and inexpensive approach for bacterial eradication that still requires scientific assessment. This study investigated new strategies for disinfection based on far-UV-C (222 nm) combined with chlorine and mechanical cleaning, providing an innovative solution using low doses. The bactericidal activity of far-UV-C (222 nm) was tested at an intensity of irradiation from 78.4 µW/cm2 to 597.7 µW/cm2 (for 1 min) against Escherichia coli and Staphylococcus epidermidis adhered on polystyrene microtiter plates. It was further tested in combination with mechanical cleaning (ultrasounds for 1 min) and free chlorine (0.1, 0.5, and 1 mg/L for 5 min). The triple combination consisting of mechanical cleaning + free chlorine (0.5 mg/L) + far-UV-C (54 mJ/cm2) was tested against cells adhered to materials found in hospital settings and other public spaces: polyvinyl chloride (PVC), stainless steel (SS), and polyetheretherketone (PEEK). Disinfection with far-UV-C (54 mJ/cm2) and free chlorine at 0.5 mg/L for 5 min allowed a total reduction of culturable E. coli cells and a logarithmic reduction of 2.98 ± 0.03 for S. epidermidis. The triple combination of far-UV-C, free chlorine, and mechanical cleaning resulted in a total reduction of culturable cells for both adhered bacteria. Bacterial adhesion to PVC, SS, and PEEK occurred at distinct extents and influenced the bactericidal activity of the triple combination, with logarithmic reductions of up to three. The overall results highlight that, based on culturability assessment, far-UV-C (54 mJ/cm2) with chlorine (0.5 mg/L; 5 min) and mechanical cleaning (1 min) as an efficient disinfection strategy using mild conditions. The combination of culturability and viability assessment of disinfection is recommended to detect regrowth events and increase the effectiveness in microbial growth control.

INVESTIGATING TRIBUTYLTIN'S TOXIC EFFECTS: INTESTINAL BARRIER AND NEUROENTERIC DISRUPTION IN RAT'S JEJUNUM.

The expansion of economic activities in coastal areas has significantly increased chemical contamination, leading to major environmental challenges. Contaminants enter the human body through the food chain, particularly via seafood and water consumption, triggering biomagnification and bioaccumulation processes. The gastrointestinal tract (GIT) acts as a selective barrier, protecting against chemical pollutants and maintaining homeostasis through a complex network of cells and immune responses. This study assessed impact of tributyltin (TBT), a highly toxic organometallic compound used in antifouling coatings for ships, on the GIT and myenteric neural plasticity in young rats. TBT exposure leads to histopathological changes, including epithelial detachment and inflammatory foci, especially at lower environmental doses. The study found that TBT causes significant reductions in villi height, increases in goblet cells and intraepithelial lymphocytes, and disrupts the myenteric plexus, with higher densities of extraganglionic neurons in exposed animals.

Mutations in ACTN4, encoding alpha-actinin-4, cause familial focal segmental glomerulosclerosis.

Focal and segmental glomerulosclerosis (FSGS) is a common, non-specific renal lesion. Although it is often secondary to other disorders, including HIV infection, obesity, hypertension and diabetes, FSGS also appears as an isolated, idiopathic condition. FSGS is characterized by increased urinary protein excretion and decreasing kidney function. Often, renal insufficiency in affected patients progresses to end-stage renal failure, a highly morbid state requiring either dialysis therapy or kidney transplantation. Here we present evidence implicating mutations in the gene encoding alpha-actinin-4 (ACTN4; ref. 2), an actin-filament crosslinking protein, as the cause of disease in three families with an autosomal dominant form of FSGS. In vitro, mutant alpha-actinin-4 binds filamentous actin (F-actin) more strongly than does wild-type alpha-actinin-4. Regulation of the actin cytoskeleton of glomerular podocytes may be altered in this group of patients. Our results have implications for understanding the role of the cytoskeleton in the pathophysiology of kidney disease and may lead to a better understanding of the genetic basis of susceptibility to kidney damage.

Milk proteins as a feed restriction signature indicating the metabolic adaptation of dairy cows.

Milk production in dairy cows is affected by numerous factors, including diet. Feed restriction is known to have little impact on milk total protein content but its effect on the fine protein composition is still poorly documented. The objective of this study was to describe the effects of two feed restriction trials of different intensities on the milk protein composition of Holstein cows. One restriction trial was of high intensity (H: 8 mid-lactation Holstein cows) and the second of moderate intensity (M: 19 peak lactation Holstein cows). Feed restriction decreased the milk protein yield for caseins under the M trial and of all six major milk proteins under the H trial. These decreased yields lead to lower concentrations of αs1-, αs2- and ß-caseins during the H trial. The milk proteome, analyzed on 32 milk samples, was affected as a function of restriction intensity. Among the 345 proteins identified eight varied under the M trial and 160 under the H trial. Ontology analyses revealed their implication in carbohydrate, lipid and protein metabolisms as well as in the immune system. These proteins reflected adaptations of the animal and mammary gland physiology to feed restriction and constituted a signature of this change.

Comparative study of bipolar eletrocoagulation versus argon plasma coagulation for rectal bleeding due to chronic radiation coloproctopathy.

BACKGROUND AND STUDY AIM: Chronic radiation coloproctopathy (CRCP) is a well-recognized complication of radiotherapy, with rectal bleeding the most common presentation. It is frequently refractory to conservative management, but the optimal endoscopic treatment of bleeding secondary to CRCP is still controversial. The efficacy and safety of bipolar eletrocoagulation (BEC) and argon plasma coagulation (APC) in the management of bleeding from CRCP were evaluated and compared. PATIENTS AND METHODS: 30 patients (mean age 67.4 years) with active and chronic bleeding from telangiectasias, were randomly allocated to BEC or APC and stratified by severity of CRCP according to clinical severity and endoscopic findings (Saunders score). Success was defined as eradication of all telangiectasias, and therapeutic failure as need for more than seven sessions or for other treatment. Complications were categorized as minor (e.g. fever, anal or abdominal pain) or major (hemorrhagic). RESULTS: Both treatments were equally effective for the treatment of CRCP rectal bleeding. Only one failure was observed in each group (P = 1.000). There was no significant difference between the two groups regarding number of sessions, minor or major complications, or relapse. However, overall complication rate was significantly higher in the BEC group (P = 0.003). CONCLUSIONS: BEC and APC are both effective for the therapy of bleeding telangiectasias from CRCP. There are probably no major differences between them. Although APC seemed safer than BEC in this investigation, further studies, involving a much larger population, are needed to assess the complication rates and determine the best management option.

The incidence of bacteraemia after argon plasma coagulation in patients with chronic radiation proctocolitis.

AIM: Argon plasma coagulation (APC) is considered a safe treatment for haemorrhagic chronic radiation proctocolitis (CRPC), but bacteraemia is a rare complication. The study aimed to evaluate the frequency of bacteraemia after APC. METHOD: A prospective study of 21 patients who underwent APC (30 procedures) for CRPC was carried out. Blood cultures (Bactec(®) ) were obtained before and 30 min after the procedure (60 samples total). Patients were monitored for 48 h after the procedure to detect signs of infection. RESULTS: None of the 21 patients had fever or any sign suggestive of infection after any of the 30 sessions. All baseline blood cultures were negative and two (7%) of the 30-min blood cultures were positive (Staphylococcus hominis n = 1; Streptococcus bovis and Rhodotorula sp n = 1). The first was likely to be a contaminant and the second patient had no evidence of any other colonic disease (neoplasia or polyps) beside CRPC. CONCLUSION: APC is a low-risk procedure regarding bacteraemia and does not warrant prophylactic antibiotic administration.

Are green roofs the path to clean air and low carbon cities?

Green roofs, as part of urban green structures, have been pointed out as the solution to pursuit the goal of healthy cities. This study aims to investigate the direct, focused on meteorological changes, and indirect, related to both meteorological and emissions changes, impacts of green roofs on air quality (PM10, NO2 and O3). For that, the numerical modelling system composed by the WRF-SLUCM-CHIMERE models was applied to a 1-year period (2017), having as case study the Porto urban area. The EnergyPlus model was also applied to estimate the green roofs impacts on the building's energy needs and related impacts on air quality and atmospheric emissions. The analysis of the direct impacts showed that green roofs promote a temperature increase during the autumn and winter seasons and a temperature decrease during the spring and summer seasons. Both negative - concentrations increase - and positive - concentrations decrease - impacts were obtained for the primary, PM10 and NO2, and secondary, O3, air pollutants, respectively, due to changes in the dynamical structure of the urban boundary layer. The indirect effects of green roofs showed their potential to enhance the buildings energy efficiency, reducing the cooling and heating needs. These changes in energy consumption promoted an overall decrease of the environmental and economic indicators. Regarding air quality, the impact was negligible. The obtained results highlight the need for a multipurpose evaluation of the impacts of green roofs, with the different effects having to be traded off against each other to better support the decision-making process.