A systematic review of the relationship between housing environmental factors and bovine respiratory disease in preweaned calves - Part 1: Ammonia, air microbial count, particulate matter and endotoxins.

Bovine respiratory disease (BRD) is one of the leading causes of mortality and morbidity in calves across diverse management systems. Despite expert opinion often citing the influence of housing environment on the level of respiratory disease in calf groups, there have been few reviews of environmental factors that predispose to BRD. This systematic review was undertaken to identify the measurable environmental variables associated with respiratory disease in housed preweaned calves. To achieve this Pubmed, CAB Direct and Scopus databases were searched. To be considered for inclusion, publications had to be fully published in English, published before 24 November, 2022 and include at least one measurable/ manipulated environmental variable and a standardized method of BRD detection. Twelve publications were included in this review. These examined a wide range of risk factors including air microbial count (four publications), air particulate matter (one publication); air endotoxins (one publication) and air ammonia (four publications). From the included publications, a statistically significant relationship to BRD was identified in 2/4 examining air microbial count, 1/1 examining air particulate matter, 1/1 examining air endotoxins and 2/4 examining air ammonia. This review indicated a paucity of evidence from the peer-review literature demonstrating a significant association between the many investigated exposure factors and BRD occurrence. An optimal environment for housed calves could not be clearly identified in this review.

A systematic review of the relationship between housing environmental factors and bovine respiratory disease in preweaned calves - Part 2: Temperature, relative humidity and bedding.

Bovine respiratory disease (BRD) is a challenge in all housed farming systems that raise calves. Farm to farm variation in BRD prevalence can be partially attributed to variation in host immunity, pathogens and housing environment. Unlike host immunity and BRD pathogens, housing environment has not been well investigated. The objective of this systematic review was to identify the measurable environmental variables associated with BRD in housed preweaned calves. Pubmed, CAB Direct and Scopus databases were searched. To be considered for inclusion publications had to be published in English, before 24 November, 2022 and include at least one measurable/ manipulated environmental variable and a standardized method of BRD detection. In total 12 publications were included in this review. In this second part of the systematic review the environmental variables identified were; temperature (9 publications); relative humidity (8 publications); bedding (5 publications); ventilation (1 publication); air CO2 concentration (1 publication) and air velocity (4 publications). Of the publications that were examined a statistically significant relationship to BRD was identified in 4/9 publications examining temperature, 3/8 examining relative humidity, 2/4 examining air velocity, 2/5 examining bedding, 0/1 examining ventilation rates and 0/1 examining CO2 concentration. From this review it is clear high airspeed at calf level should be avoided as should deep, wet pack bedding. The relationship between BRD prevalence and both high and low temperature requires more investigation to identify temperature thresholds associated with increased risk of BRD as well as the most influential modifiers. An optimal environment for housed calves could not be clearly identified in this review.

Obesity and Hyperandrogenemia in Polycystic Ovary Syndrome: Clinical Implications.

Polycystic ovary syndrome (PCOS) is often accompanied with metabolic disturbances attributed to androgen excess and obesity, but the contribution of each has not been defined, and the occurrence of metabolic disturbances is usually not investigated. Ninety-nine women with PCOS and forty-one without PCOS were evaluated. The clinical biomarkers of alterations related to glucose (glucose, insulin, and clamp-derived glucose disposal - M), liver (aspartate aminotransferase, alanine aminotransferase, and gamma-glutamyl transferase), and endothelium (arginine, asymmetric dymethylarginine, carotid intima-media thickness, and flow-mediated dilation) metabolism were measured; participants were categorized into four groups according to their obesity (OB) and hyperandrogenemia (HA) status as follows: Healthy (no-HA, lean), HA (HA, lean), OB (no-HA, OB), and HAOB (HA, OB). Metabolic disturbances were very frequent in women with PCOS (≈70%). BMI correlated with all biomarkers, whereas free testosterone (FT) correlated with only glucose- and liver-related indicators. Although insulin sensitivity and liver enzymes were associated with FT, women with obesity showed lower M (coef = 8.56 - 0.080(FT) - 3.71(Ob); p < 0.001) and higher aspartate aminotransferase (coef = 26.27 + 0.532 (FT) + 8.08 (Ob); p = 0.015) than lean women with the same level of FT. Women with obesity showed a higher risk of metabolic disorders than lean women, independent of hyperandrogenemia. Clinicians are compelled to look for metabolic alterations in women with PCOS. Obesity should be treated in all cases, but hyperandrogenemia should also be monitored in those with glucose-or liver-related disturbances.

Adenovirus type D and type E infection in broiler chickens: the effect on CD4 and CD8 T cell response, cytokines expression and their immunopathology.

1. A total of 150-day-old chicks were divided into three groups of 50 birds (G1-G3); G1 and G2 were orally inoculated at 1-day old with 0.5 ml of 107 TCID50/ml FAdV-D serotype 2 (MT386509.1) and FAdV-E serotype 8a (MW847902), respectively, and G3 was blank control group.2. Cell-mediated immune response was evaluated by detection of CD4, CD8 T lymphocytes and the mRNA expression of IL6 and IL8 in the chicken spleen using q-PCR. Additionally, immunopathology was performed at 3, 5 and 7 day post infection (dpi) and weekly until the end of the experiment.3. Results revealed that transcription of inflammatory cytokines (IL6, IL8) was up regulated in the spleen of FAdV type D and type E infected chickens at various time points relative to the control group. A marked decrease in the number of CD4 and CD8 T lymphocytes at 5 and 7 dpi in G1 of chickens infected with FAdV type D. Whereas, in chickens infected with FAdV type E, the CD4 and CD8 T lymphocytes were markedly decreased at 7 dpi.4. In contrast, there were no significant differences in humoral immune responses against NDV vaccine in (G1 and G2) at different intervals post-vaccination compared to the control group. The histopathology of the bursa, thymus, and spleen in the infected groups showed lymphocytolysis with severe reticular cells hyperplasia and lymphoid depletion.5. In conclusion, fowl adenovirus types D and E have an immunosuppressive effect in broilers which may be considered one of the main causes of the continuous co-infections with other viruses reported in the field during the last 10 years.

Stability of a delayed SARS-CoV-2 reactivation model with logistic growth and adaptive immune response.

This paper develops and analyzes a SARS-CoV-2 dynamics model with logistic growth of healthy epithelial cells, CTL immune and humoral (antibody) immune responses. The model is incorporated with four mixed (distributed/discrete) time delays, delay in the formation of latent infected epithelial cells, delay in the formation of active infected epithelial cells, delay in the activation of latent infected epithelial cells, and maturation delay of new SARS-CoV-2 particles. We establish that the model's solutions are non-negative and ultimately bounded. We deduce that the model has five steady states and their existence and stability are perfectly determined by four threshold parameters. We study the global stability of the model's steady states using Lyapunov method. The analytical results are enhanced by numerical simulations. The impact of intracellular time delays on the dynamical behavior of the SARS-CoV-2 is addressed. We noted that increasing the time delay period can suppress the viral replication and control the infection. This could be helpful to create new drugs that extend the delay time period.

Ecological Calendars, Food Sovereignty, and Climate Adaptation in Standing Rock.

Indigenous food sovereignty relies on ecological knowledge of plants and animals, including knowledge related to their development and behavior through the seasons. In the context of anthropogenic climate change, ecological calendars based on Indigenous knowledge may enable communities to anticipate seasonal phenomena. We conducted research with communities in the Standing Rock Nation (North and South Dakota, USA) to develop ecological calendars based on their ecological knowledge. We present ecological calendars developed in seven communities through a series of workshops and interviews. These calendars are rich with knowledge about temporal relations within each community's ecosystem, including the use of plants and animals as seasonal indicators and cues for food system activities. However, the calendars also reveal the impacts of cultural genocide wrought by the United States government in its efforts to colonize the lands and minds of Indigenous communities. Given the diversity of knowledge among Standing Rock communities, we identify opportunities for knowledge exchange to revitalize ecological relations at the heart of food sovereignty. We highlight the potential for ecological calendars to facilitate climate adaptation by enabling communities to synchronize their food systems with an increasingly variable climate.

Leptin receptor and prolactin in pubertal disorders and chronic kidney disease.

BACKGROUND: Knowledge of chronic kidney disease (CKD) with pubertal disorders (PD) in adolescent boys is limited as few studies have explored this disorder. This study aimed to identify the usefulness of assessing hormonal parameters in male adolescents with CKD and their correlation with PD in a 12-month follow-up period. METHODS: A prospective cohort study was conducted among male adolescents with CKD (stages IV and V). Data regarding the age at puberty onset were collected from the patients' clinical records and through interview. The patients were followed up for 12 months during their pubertal development. At the beginning, routine hormonal profile tests were performed to examine the patients' thyroid profile, prolactin levels, luteinizing hormone, follicle-stimulating hormone, testosterone, leptin, and receptor leptin. The hormonal profiles of patients with and without PD were compared. Comparisons between the groups were performed using the Student t-test and Fisher's exact tests. Logistic regression analysis was also performed. RESULTS: Data of 64 patients (26/64 with PD) were analyzed. The median age was 15 years and the median time for CKD evolution was 11 months. No differences between groups were noted in the general or biochemical characteristics of the patients. The hormonal parameters, prolactin levels were higher and the free leptin and free thyroxine levels were lower in patients with PD. Leptin receptor levels of >0.90 ng/mL (risk ratio [RR], 8.6; P = 0.004) and hyperprolactinemia (RR, 21.3; P = 0.049) were the risk factors for PD. CONCLUSIONS: Leptin receptor levels of >0.90 ng/mL and hyperprolactinemia are associated with the development of PD in male adolescents with CKD.

Role of Biodiversity in Ecological Calendars and Its Implications for Food Sovereignty: Empirical Assessment of the Resilience of Indicator Species to Anthropogenic Climate Change.

Ecological calendars are knowledge systems based on close observation of one's habitat to measure and give meaning to time, thereby providing anticipatory capacity for livelihood activities and contributing to food sovereignty. They rely upon seasonal indicators that integrate biophysical and ecological phenomena (e.g., first snowfall, last frost, blossoming of a tree species; seasonal appearance of an animal or plant) with locally grounded cultural meaning and value systems. These context-specific relationships have enabled Indigenous and rural societies to anticipate weather and other seasonal processes in their environment. However, anthropogenic climate change could undermine ecological calendars due to adverse impacts on specific indicators species, but this issue remains unexplored. We address this knowledge gap by examining how anthropogenic climate change might affect selected species (birds, fish, and mammals) that are seasonal and key to Indigenous food systems in two Western Arctic communities. We leverage existing dietary animal datasets to which we apply a novel methodology for assessing organismal vulnerability to climate change. The methodology uses intrinsic species traits such as physiological tolerances, genetic variability, and life history traits to generate an empirical and integrative assessment of vulnerability for any given species. Subsequently, an aggregate view of vulnerability across calendar species is achieved through comparative statistical analysis across species both within and between communities. This exercise permits the first quantitative assessment of the continued relevance and effective use of an ecological calendar, thus demonstrating that food sovereignty and livelihood security is enhanced by biodiversity of indicator species.

Antifungal Activity of Copper Oxide Nanoparticles against Root Rot Disease in Cucumber.

Metal oxide nanoparticles have recently garnered interest as potentially valuable substances for the management of plant diseases. Copper oxide nanoparticles (Cu2ONPs) were chemically fabricated to control root rot disease in cucumbers. A scanning electron microscope (SEM), X-ray diffraction (XRD) and photoluminescence (PL) were employed to characterize the produced nanoparticles. Moreover, the direct antifungal activity of Cu2ONPs against Fusarium solani under laboratory, greenhouse, and field conditions were also evaluated. In addition, the induction of host-plant resistance by Cu2ONPs was confirmed by the results of enzyme activities (catalase, peroxidase, and polyphenoloxidase) and gene expression (PR-1 and LOX-1). Finally, the effect of Cu2ONPs on the growth and productivity characteristics of the treated cucumber plants was investigated. The average particle size from all the peaks was found to be around 25.54 and 25.83 nm for 0.30 and 0.35 Cu2O, respectively. Under laboratory conditions, the study found that Cu2ONPs had a greater inhibitory effect on the growth of Fusarium solani than the untreated control. Cu2ONP treatment considerably reduced the disease incidence of the root rot pathogen in cucumber plants in both greenhouse and field environments. Defense enzyme activity and defense genes (PR1 and LOX1) transcription levels were higher in cucumber plants treated with Cu2ONPs and fungicide than in the untreated control. SEM analysis revealed irregularities, changes, twisting, and plasmolysis in the mycelia, as well as spore shrinking and collapsing in F. solani treated with Cu2ONPs, compared to the untreated control. The anatomical analysis revealed that cucumber plants treated with Cu2ONPs had thicker cell walls, root cortex, and mesophyll tissue (MT) than untreated plants. Cucumber growth and yield characteristics were greatly improved after treatment with Cu2ONPs and fungicide. To the best of our knowledge, employing Cu2ONPs to treat cucumber rot root disease is a novel strategy that has not yet been reported.

First test-day postcalving risk factors for clinical mastitis in southern Chile dairy farms: A retrospective cohort study.

We conducted a retrospective cohort study to evaluate risk factors associated with clinical mastitis in dairy cows in southern Chile between the first milk test after calving to 200 d in milk (DIM). Data from 8,772 cows were collected from June 2017 to November 2019 on 10 dairy farms selected by convenience in southern Chile. Data were analyzed using a multivariable mixed logistic regression using a manual backward selection process with logit link function and farm as a random effect. The dependent variable was clinical mastitis between the first milking sample collected by monthly milk sampling program up to 200 DIM. Days in milk, fat, urea, parity, season, and somatic cell counts were statistically associated with clinical mastitis in the first 200 d in milk. Cows with higher than average milk yield, parity, urea, and somatic cell count were at greater odds of being diagnosed with clinical mastitis compared with their respective referents. In contrast, higher milk fat was associated with lower odds of clinical mastitis. Significant interactions between days in milk with season and parity were observed, where for every 5-d increase in DIM, the odds of clinical mastitis decreased by different proportions depending on the interaction. Identified risk factors for clinical mastitis using first monthly milk sampling data can help dairy farmers in Chile implement herd-level mastitis prevention and control measures.

The socioeconomic impact of multiple sclerosis in France: Results from the PETALS study.

Background: Multiple sclerosis (MS) places a considerable financial burden on the society. However, data quantifying the contemporary cost burden in France are lacking. Objective: This cost-of-illness study aimed to estimate the direct and indirect costs associated with MS in France. Methods: Between October 2020-November 2020, 208 French adults with a confirmed diagnosis of MS were recruited via MSCopilot® (a new MS self-assessment digital solution) and several MS patient networks. Indirect costs were estimated using a combination of top-down and bottom-up approaches. Direct costs were retrieved from Assurance Maladie (i.e. national system of health insurance) publications. Out-of-pocket expenses (OOPEs) incurred by MS patients were also reported. All costs were expressed in €2020. Data from the survey were extrapolated to the overall French MS population. Results: MS exerted an annual cost burden of €2.7 billion on the French society (indirect costs: €1.3 billion; direct costs: €1.4 billion). Mean annual costs were €27,164.7 per-patient, with indirect and direct costs accounting for 48.1% and 51.9% of the total annual costs, respectively. OOPEs contributed over €90 million to the total annual costs. Conclusions: MS imposes a substantial cost burden on the French society, with approximately half of the total annual costs driven by indirect costs.

Efficacy and Safety of Vitamin D Supplementation to Prevent COVID-19 in Frontline Healthcare Workers. A Randomized Clinical Trial.

BACKGROUND: Associations between vitamin D (VD) deficiency and the risk of SARS-CoV-2 infection have been documented in cross-sectional population studies. Intervention studies in patients with moderate to severe COVID-19 have failed to consistently document a beneficial effect. OBJECTIVE: To determine the efficacy and safety of VD-supplementation in the prevention of SARS-CoV-2 infection in highly exposed individuals. METHODS: A double-blind, parallel, randomized trial was conducted. Frontline healthcare workers from four hospitals in Mexico City, who tested negative for SARS-CoV-2 infection, were enrolled between July 15 and December 30, 2020. Participants were randomly assigned to receive 4,000 IU VD (VDG) or placebo (PG) daily for 30 d. RT-PCR tests were taken at baseline and repeated if COVID-19 manifestations appeared during follow-up. Serum 25-hydroxyvitamin D3 and antibody tests were measured at baseline and at day 45. Per-protocol and intention-to-treat analysis were conducted. RESULTS: Of 321 recruited subjects, 94 VDG and 98 PG completed follow-up. SARS-CoV-2 infection rate was lower in VDG than in PG (6.4 vs. 24.5%, p <0.001). The risk of acquiring SARS-CoV-2 infection was lower in the VDG than in the PG (RR: 0.23; 95% CI: 0.09-0.55) and was associated with an increment in serum levels of 25-hydroxyvitamin D3 (RR: 0.87; 95% CI: 0.82-0.93), independently of VD deficiency. No significant adverse events were identified. CONCLUSIONS: Our results suggest that VD-supplementation in highly exposed individuals prevents SARS-CoV-2 infection without serious AEs and regardless of VD status.

Enteric protists in wild western chimpanzees (Pan troglodytes verus) and humans in Comoé National Park, Côte d'Ivoire.

The western chimpanzee (Pan troglodytes verus), a subspecies of the common chimpanzee, is currently listed as Critically Endangered. Human-driven habitat loss and infectious diseases are causing dramatic chimpanzee population declines and range contractions that are bringing these primates to the brink of extinction. Little information is currently available on the occurrence of diarrhoea-causing enteric protist species in chimpanzees in general, and in western chimpanzees in particular, or on the role of humans as a potential source of these infections. In this prospective molecular epidemiological study, we investigated the presence, genetic variability, and zoonotic potential of enteric protists in faecal samples from western chimpanzees (n = 124) and humans (n = 9) in Comoé National Park, Côte d'Ivoire. Parasite detection and genotyping were conducted by using polymerase chain reaction (PCR) and Sanger sequencing. The protist species found in the chimpanzee samples were Entamoeba dispar (14.5%), Blastocystis sp. (11.3%), Giardia duodenalis (5.8%), Troglodytella abrassarti (2.5%) and Cryptosporidium hominis (0.8%). The protist species found in the human samples were G. duodenalis (22.2%) and Blastocystis sp. (11.1%). Entamoeba histolytica, Enterocytozoon bieneusi, and Balantioides coli were undetected in both chimpanzee and human samples. Sequence analyses revealed the presence of Blastocystis subtype (ST) 1 (alleles 4 and 8) and ST3 (allele 24) in chimpanzees, and ST3 (allele 52) in humans. ST1 allele 8 represents a chimpanzee-adapted Blastocystis genetic variant. Cross-species transmission of pathogenic enteric protists between chimpanzees and humans might be possible in Comoé National Park, although the frequency and extent of zoonotic events remain to be fully elucidated.

Presence and genetic diversity of enteric protists in captive and semi-captive non-human primates in côte d'Ivoire, Sierra Leone, and Peru.

Little information is currently available on the occurrence and genetic diversity of pathogenic and commensal protist species in captive and semi-captive non-human primates (NHP) resident in zoological gardens or sanctuaries in low- and medium-income countries. In this molecular-based study, we prospectively collected individual faecal samples from apparently healthy NHP at the Abidjan Zoological Garden (AZG) in Côte d'Ivoire, the Tacugama Sanctuary (TS) in Sierra Leone, and the Quistococha Zoological Garden (QZG) in Peru between November 2018 and February 2020. We evaluated for the presence of pathogenic (Cryptosporidium spp., Entamoeba histolytica, Giardia duodenalis, Blastocystis sp., Enterocytozoon bieneusi, Balantioides coli) and commensal (Entamoeba dispar, Troglodytella abrassarti) protist species using PCR methods and Sanger sequencing. Giardia duodenalis was the most prevalent species found (25.9%, 30/116), followed by Blastocystis sp. (22.4%, 26/116), and E. dispar (18.1%, 21/116). We detected E. bieneusi (4.2%, 1/24) and T. abrassarti (12.5%, 3/24) only on NHP from AZG. Cryptosporidium spp., E. histolytica, and B. coli were undetected at the three sampling sites investigated here. Sequence analyses revealed the presence of zoonotic sub-assemblages BIII (n = 1) in AZG and BIV (n = 1) in TS within G. duodenalis. We identified Blastocystis subtype ST3 (100%, 6/6) in AZG, ST1 (80.0%, 12/15), ST2 (6.7%, 1/15), and ST3 (13.3%, 2/15) in TS, and ST2 (80.0%, 4/5) and ST3 (20.0%, 1/5) in QZG. The only E. bieneusi isolate detected here was identified as zoonotic genotype CAF4. Our PCR-based data indicate that potentially pathogenic protist species including G. duodenalis, Blastocystis sp., E. bieneusi, and B. coli are present at variable rates in the three NHP populations investigated here. The identification of zoonotic genotypes within these species indicates that human-NHP transmission is possible, although the extent and directionality of these events need to be elucidated in future molecular surveys.

Validation of Cardiac Surgery-Associated Neutrophil Gelatinase-Associated Lipocalin Score for Prediction of Cardiac Surgery-Associated Acute Kidney Injury.

BACKGROUND: The Cardiac Surgery-Associated Neutrophil Gelatinase-Associated Lipocalin (CSA-NGAL) score has been developed to stratify patients with cardiac surgery-associated acute kidney injury (CSA-AKI). Its predictive power needs to be validated to guide clinical decision for such high-risk patients. METHODS: A prospective study was conducted on 637 consecutive adult patients who developed postoperative AKI after cardiac surgery with cardiopulmonary bypass. AKI was defined according to Kidney Disease: Improving Global Outcomes criteria (KDIGO). The CSA-NGAL score was calculated. Assessment of the diagnostic performance of the scoring model was performed by area under the receiver operating curve analysis. RESULTS: The area under the curve for the postoperative Urinary NGAL showed an area under the curve ([standard error (SE)] 0.80 (0.38); p<0.001; 95% CI 0.72-0.87). Its sensitivity for CSA-AKI in the first 24 hours was 66% and specificity was 80% (cut-off value 300.1 ng/mL). There was a positive correlation between NGAL score and KDIGO criteria, with a significant increase in postoperative mean Urinary NGAL values as the KDIGO stage increased. CONCLUSION: The CSA-NGAL score has a high sensitivity, specificity and positive predictive value that can translate into improved outcomes and resource allocation. It is believed that adding it to the existing clinical scoring systems for AKI prediction will be productive.

Molecular Detection and Characterization of Intestinal and Blood Parasites in Wild Chimpanzees (Pan troglodytes verus) in Senegal.

Wild chimpanzee populations in West Africa (Pan troglodytes verus) have dramatically decreased as a direct consequence of anthropogenic activities and infectious diseases. Little information is currently available on the epidemiology, pathogenic significance, and zoonotic potential of protist species in wild chimpanzees. This study investigates the occurrence and genetic diversity of intestinal and blood protists as well as filariae in faecal samples (n = 234) from wild chimpanzees in the Dindefelo Community Nature Reserve, Senegal. PCR-based results revealed the presence of intestinal potential pathogens (Sarcocystis spp.: 11.5%; Giardia duodenalis: 2.1%; Cryptosporidium hominis: 0.9%), protist of uncertain pathogenicity (Blastocystis sp.: 5.6%), and commensal species (Entamoeba dispar: 18.4%; Troglodytella abrassarti: 5.6%). Entamoeba histolytica, Enterocytozoon bieneusi, and Balantioides coli were undetected. Blood protists including Plasmodium malariae (0.4%), Trypanosoma brucei (1.3%), and Mansonella perstans (9.8%) were also identified. Sanger sequencing analyses revealed host-adapted genetic variants within Blastocystis, but other parasitic pathogens (C. hominis, P. malariae, T. brucei, M. perstans) have zoonotic potential, suggesting that cross-species transmission between wild chimpanzees and humans is possible in areas where both species overlap. Additionally, we explored potential interactions between intestinal/blood protist species and seasonality and climate variables. Chimpanzees seem to play a more complex role on the epidemiology of pathogenic and commensal protist and nematode species than initially anticipated.

Molecular Detection and Characterization of Blastocystis sp. and Enterocytozoon bieneusi in Cattle in Northern Spain.

Some enteric parasites causing zoonotic diseases in livestock have been poorly studied or even neglected. This is the case in stramenopile Blastocystis sp. and the microsporidia Enterocytozoon bieneusi in Spain. This transversal molecular epidemiological survey aims to estimate the prevalence and molecular diversity of Blastocystis sp. and E. bieneusi in cattle faecal samples (n = 336) in the province of Álava, Northern Spain. Initial detection of Blastocystis and E. bieneusi was carried out by polymerase chain reaction (PCR) and Sanger sequencing of the small subunit (ssu) rRNA gene and internal transcribed spacer (ITS) region, respectively. Intra-host Blastocystis subtype diversity was further investigated by next generation amplicon sequencing (NGS) of the ssu rRNA gene in those samples that tested positive by conventional PCR. Amplicons compatible with Blastocystis sp. and E. bieneusi were observed in 32.1% (108/336, 95% CI: 27.2-37.4%) and 0.6% (2/336, 95% CI: 0.0-1.4%) of the cattle faecal samples examined, respectively. Sanger sequencing produced ambiguous/unreadable sequence data for most of the Blastocystis isolates sequenced. NGS allowed the identification of 10 Blastocystis subtypes including ST1, ST3, ST5, ST10, ST14, ST21, ST23, ST24, ST25, and ST26. All Blastocystis-positive isolates involved mixed infections of 2-8 STs in a total of 31 different combinations. The two E. bieneusi sequences were confirmed as potentially zoonotic genotype BEB4. Our data demonstrate that Blastocystis mixed subtype infections are extremely frequent in cattle in the study area. NGS was particularly suited to discern underrepresented subtypes or mixed subtype infections that were undetectable or unreadable by Sanger sequencing. The presence of zoonotic Blastocystis ST1, ST3, and ST5, and E. bieneusi BEB4 suggest cross-species transmission and a potential risk of human infection/colonization.

Does the Nuss procedure for treating pectus excavatum compromise the long term internal mammary artery flow? A systematic review.

INTRODUCTION: The Nuss procedure is the most common procedure used to treat patients with pectus excavatum. The effect of the Nuss bars on the long-term internal mammary artery flow (IMA) is not well studied. This could have an impact on patients requiring a coronary artery bypass grafting surgery after the Nuss procedure. We performed a systematic review to study the impact of the Nuss bars on the IMA long term flow. METHODS: A Medline search from January 1990 to August 2020 was performed using [Nuss OR thoracoscopic pectus OR minimally invasive pectus] AND [Internal mammary OR Internal thoracic OR IMA OR ITA]. English language papers only were included. This trial was registered with PROSPERO under registration number CRD42021234010. RESULTS: A total of 48 papers were identified using the reported search, of which three represented the best evidence to answer the clinical question. One study looked at the IMA flow via computed tomography (CT)-angiography on the 10th postoperative day after the Nuss procedure and found 15 out of 34 patients (44%) to have abnormal IMA blood flow but with no clinical consequences. Two studies looked at the IMA flow after removal of the Nuss-bar. The first study utilized CT-angiography on the 5th postoperative day after Nuss-bar removal and found four out of the six patients studied (67%) to have abnormal flow. The last study was composed of 19 patients and looked at IMA flow during the presence of the Nuss-bars and after its removal utilizing Doppler-angiography. It found 11 out of 19 patients (58%) to have abnormal blood flow with the bars in place. After removal of the bars, only two patients (10%) were found to have unilateral IMA obstructed flow. CONCLUSION: In patients undergoing the Nuss procedure for management of pectus excavatum, the internal mammary artery flow is compromised in 44%-58% of patients with the bar in situ. When these patients are assessed 10 days following removal of the bar, some reversal of compromised IMA flow is evident. However, in up to 67% of patients, abnormal IMA flow remains. Further studies are required to determine whether this abnormal flow is permanent, which will require examining patients at longer follow-up intervals. Patients undergoing coronary artery bypass grafting who have a history of a Nuss procedure should receive preoperative IMA imaging.

Occurrence and Genetic Diversity of Protist Parasites in Captive Non-Human Primates, Zookeepers, and Free-Living Sympatric Rats in the Córdoba Zoo Conservation Centre, Southern Spain.

Little information is currently available on the epidemiology of parasitic and commensal protist species in captive non-human primates (NHP) and their zoonotic potential. This study investigates the occurrence, molecular diversity, and potential transmission dynamics of parasitic and commensal protist species in a zoological garden in southern Spain. The prevalence and genotypes of the main enteric protist species were investigated in faecal samples from NHP (n = 51), zookeepers (n = 19) and free-living rats (n = 64) by molecular (PCR and sequencing) methods between 2018 and 2019. The presence of Leishmania spp. was also investigated in tissues from sympatric rats using PCR. Blastocystis sp. (45.1%), Entamoeba dispar (27.5%), Giardia duodenalis (21.6%), Balantioides coli (3.9%), and Enterocytozoon bieneusi (2.0%) (but not Troglodytella spp.) were detected in NHP. Giardia duodenalis (10.5%) and Blastocystis sp. (10.5%) were identified in zookeepers, while Cryptosporidium spp. (45.3%), G. duodenalis (14.1%), and Blastocystis sp. (6.25%) (but not Leishmania spp.) were detected in rats. Blastocystis ST1, ST3, and ST8 and G. duodenalis sub-assemblage AII were identified in NHP, and Blastocystis ST1 in zookeepers. Giardia duodenalis isolates failed to be genotyped in human samples. In rats, four Cryptosporidium (C. muris, C. ratti, and rat genotypes IV and V), one G. duodenalis (assemblage G), and three Blastocystis (ST4) genetic variants were detected. Our results indicate high exposure of NHP to zoonotic protist species. Zoonotic transmission of Blastocysts ST1 was highly suspected between captive NHP and zookeepers.

Molecular Diversity of Giardia duodenalis, Cryptosporidium spp., and Blastocystis sp. in Symptomatic and Asymptomatic Schoolchildren in Zambézia Province (Mozambique).

Infections by the protist enteroparasites Giardia duodenalis, Cryptosporidium spp., and, to a much lesser extent, Blastocystis sp. are common causes of childhood diarrhoea in low-income countries. This molecular epidemiological study assesses the frequency and molecular diversity of these pathogens in faecal samples from asymptomatic schoolchildren (n = 807) and symptomatic children seeking medical attention (n = 286) in Zambézia province, Mozambique. Detection and molecular characterisation of pathogens was conducted by polymerase chain reaction (PCR)-based methods coupled with Sanger sequencing. Giardia duodenalis was the most prevalent enteric parasite found [41.7%, 95% confidence interval (CI): 38.8‒44.7%], followed by Blastocystis sp. (14.1%, 95% CI: 12.1‒16.3%), and Cryptosporidium spp. (1.6%, 95% CI: 0.9‒2.5%). Sequence analyses revealed the presence of assemblages A (7.0%, 3/43) and B (88.4%, 38/43) within G. duodenalis-positive children. Four Cryptosporidium species were detected, including C. hominis (30.8%; 4/13), C. parvum (30.8%, 4/13), C. felis (30.8%, 4/13), and C. viatorum (7.6%, 1/13). Four Blastocystis subtypes were also identified including ST1 (22.7%; 35/154), ST2 (22.7%; 35/154), ST3 (45.5%; 70/154), and ST4 (9.1%; 14/154). Most of the genotyped samples were from asymptomatic children. This is the first report of C. viatorum and Blastocystis ST4 in Mozambique. Molecular data indicate that anthropic and zoonotic transmission (the latter at an unknown rate) are important spread pathways of diarrhoea-causing pathogens in Mozambique.