Association of Serum Ferritin Levels with Microalbuminuria, Glycemic Control and Dyslipidemia.

As diabetes is on the rise worldwide, early screening and detection of its complications is crucial. The first clinical sign of renal dysfunction in diabetes is microalbuminuria. Ferritin may have a possible role in the development and prediction of microalbuminuria and other complications of diabetes by 3 possible mechanisms- 1) it is a marker of elevated body iron stores. 2) acts as acute-phase reactant and elevation reflects inflammation 3) Delayed clearance of glycosylated ferritin in patients with diabetes causes raised ferritin levels. In this premise, we studied the association between serum ferritin levels and microalbuminuria, glycemic control and dyslipidemia in type 2 diabetic patients. MATERIAL: A cross-sectional study was carried out at a tertiary hospital in northern India involving 152 patients of type 2 diabetes mellitus. Anthropometric measurements (weight, height), sample collection for basic laboratory parameters including complete blood count, kidney function tests, liver function tests, HbA1c, lipid profile, serum iron and ferritin levels, urinary albumin- creatinine ratio was done. A fundus examination was performed. Results were analyzed using descriptive statistics and making comparisons among various groups. Categorical data were summarized as proportions or percentages while discrete as mean. Chi-square test was used for testing associations. OBSERVATION: Mean serum ferritin levels were significantly higher in patients having microalbuminuria as against patients having normal urinary albumin- creatinine ratio (539 vs 292, p-value- 0.04). Patients having poor glycemic control (HbA1c > 7) had higher ferritin levels (p-value- 0.024). There was a near-significant association (p-value-0.06) between serum ferritin and triglyceride levels. More cases with raised serum ferritin had higher triglyceride levels. Patients having microalbuminuria had a higher prevalence of diabetic retinopathy (p-value- 0.003) and higher triglyceride levels (p-value- 0.041). CONCLUSION: Ferritin can act as a marker of diabetic nephropathy, poor glycemic control and dyslipidemia in type 2 diabetic patients. Our study, therefore, indicates that programs to prevent complications of diabetes would be more effective by placing greater emphasis on patients having higher ferritin levels.

Altered electrochemical properties of iron oxide nanoparticles by carbon enhance molecular biocompatibility through discrepant atomic interaction.

Recent advancement in nanotechnology seeks exploration of new techniques for improvement in the molecular, chemical, and biological properties of nanoparticles. In this study, carbon modification of octahedral-shaped magnetic nanoparticles (MNPs) was done using two-step chemical processes with sucrose as a carbon source for improvement in their electrochemical application and higher molecular biocompatibility. X-ray diffraction analysis and electron microscopy confirmed the alteration in single-phase octahedral morphology and carbon attachment in Fe3O4 structure. The magnetization saturation and BET surface area for Fe3O4, Fe3O4/C, and α-Fe2O3/C were measured as 90, 86, and 27 emu/g and 16, 56, and 89 m2/g with an average pore size less than 7 nm. Cyclic voltammogram and galvanostatic charge/discharge studies showed the highest specific capacitance of carbon-modified Fe3O4 and α-Fe2O3 as 213 F/g and 192 F/g. The in vivo biological effect of altered physicochemical properties of Fe3O4 and α-Fe2O3 was assessed at the cellular and molecular level with embryonic zebrafish. Mechanistic in vivo toxicity analysis showed a reduction in oxidative stress in carbon-modified α-Fe2O3 exposed zebrafish embryos compared to Fe3O4 due to despaired influential atomic interaction with sod1 protein along with significant less morphological abnormalities and apoptosis. The study provided insight into improving the characteristic of MNPs for electrochemical application and higher biological biocompatibility.

GAMOGONY OF SARCOCYSTIS STRIXI IN MAMMALIAN CELL CULTURES.

We are interested in the disease ecology of Sarcocystis species that infect birds of prey as definitive and intermediate hosts. The present study was done to test our hypothesis that a laboratory model can be developed for sarcocystis infection in mammals using gamma interferon gene knockout (KO) mice as a source of Sarcocystis strixi bradyzoites and mammalian cell cultures as a source of sporulated S. strixi oocysts. Sporocysts of S. strixi from a naturally infected barred owl (Strix varia) were fed to KO mice to produce sarcocysts, and the enclosed bradyzoites were obtained by acid-pepsin digestion of abdominal and thigh muscles. Bradyzoites, metrocytes, and an unusual spherical stage were seen in digest before the inoculation of host cells. The spherical stages stained dark with Giemsa stain, but no nucleus was observed, and they were seen free and associated with the concave portion of some bradyzoites. Examination of infected cell cultures demonstrated that macrogamonts and microgamonts were present at 24 hr post-inoculation. Since sporulated oocysts were not observed, we had to reject our current hypothesis.

Endophyte roles in nutrient acquisition, root system architecture development and oxidative stress tolerance.

Plants associate with communities of microbes (bacteria and fungi) that play critical roles in plant development, nutrient acquisition and oxidative stress tolerance. The major share of plant microbiota is endophytes which inhabit plant tissues and help them in various capacities. In this article, we have reviewed what is presently known with regard to how endophytic microbes interact with plants to modulate root development, branching, root hair formation and their implications in overall plant development. Endophytic microbes link the interactions of plants, rhizospheric microbes and soil to promote nutrient solubilization and further vectoring these nutrients to the plant roots making the soil-plant-microbe continuum. Further, plant roots internalize microbes and oxidatively extract nutrients from microbes in the rhizophagy cycle. The oxidative interactions between endophytes and plants result in the acquisition of nutrients by plants and are also instrumental in oxidative stress tolerance of plants. It is evident that plants actively cultivate microbes internally, on surfaces and in soils to acquire nutrients, modulate development and improve health. Understanding this continuum could be of greater significance in connecting endophytes with the hidden half of the plant that can also be harnessed in applied terms to enhance nutrient acquisition through the development of favourable root system architecture for sustainable production under stress conditions.

White-tailed deer (Odocoileus virginianus) are a reservoir of a diversity of Toxoplasma gondii strains in the USA and pose a risk to consumers of undercooked venison.

To assess the role of white-tailed deer (Odocoileus virginianus, WTD) in the epidemiology of toxoplasmosis, we conducted a national survey of WTD across the USA for Toxoplasma gondii infection. To do this, we combined serology with parasite isolation to evaluate the prevalence and genetic diversity of T. gondii in this game species. From October 2012 to March 2019, serum and tissues were collected from 914 WTD across the USA. Serum samples were screened for antibodies to T. gondii, and then the tissues of seropositive WTD were bioassayed in mice. Antibodies were detected in 329 (36%) of 914 WTD tested by the modified agglutination test (positive reaction at 1:25 or higher). Viable T. gondii was isolated from the heart of 36 WTD from 11 states. Three of the 36 isolates were pathogenic but not highly virulent to outbred Swiss Webster mice and all 36 isolates could be propagated further in cell culture and were genotyped. For genotyping, DNA extracted from cell culture-derived tachyzoites was characterized by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) using the genetic markers SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1 and Apico. Genotyping revealed seven ToxoDB PCR-RFLP genotypes, including 24 isolates for genotype #5 (haplogroup 12), four isolates for #2 (type III, haplogroup 3), three isolates for genotypes #1 (type II, haplogroup 2), two isolates for genotypes #3 (type II, haplogroup 2) and one isolate each for #39, #221 and #224. Genotype #5 was the most frequently isolated, accounting for 66.6% (24 of 36) of the isolates. Combining the 36 isolates from this study with previously reported 69 isolates from WTD, 15 genotypes have been identified. Among these, 50.4% (53/105) isolates belong to genotype #5. Our results indicate moderate genetic diversity of T. gondii in WTD. The results also indicate that undercooked venison should not be consumed by humans or fed to cats.

Aspergillus sensitization in bronchial asthma: A separate phenotype.

Background: Bronchial asthma is one of the common chronic respiratory illnesses worldwide, with a global disease burden that affects approximately 300 million individuals. It affects 5-10% of the population in developed countries. India constitutes approximately one-tenth of the global disease burden, where an estimated 7 million children are affected by this illness. Airway hyperresponsiveness or bronchial hyperreactivity in asthma is an exaggerated response to numerous exogenous and endogenous stimuli. In India, the most important endogenous stimuli is considered to be fungal antigens from the genus, Aspergillus. Positive skin reactions to Aspergillus fumigatus among individuals with atopic asthma is 10 to 20%; however, a causal relationship is yet to be established. A separate entity, termed "severe asthma with fungal sensitivity," is often used to describe milder allergic reactions to fungal aeroallergens that has fungal sensitization as the starting point of pathogenesis than allergic bronchopulmonary aspergillosis. Methods: A total of 57 patients with bronchial asthma were prospectively enrolled and evaluated for Aspergillus sensitization and its impact on asthma control. Results: Symptoms were well controlled in a majority of patients (57.89%) and partly controlled in 14.04%. Uncontrolled symptoms were observed in only 28.07% of patients. The proportion of patients with uncontrolled symptoms was higher among patients who were sensitized (45.00%) compared with patients who were not sensitized (18.92%). The proportion of patients with well-controlled symptoms was higher in individuals who were not sensitized compared with individuals who were sensitized (62.16% versus 50.00%). Conclusion: The severity of asthma was associated with fungal sensitization, with sensitization to Aspergillus as a part of the pathogenesis. Aspergillus sensitization is significantly associated with bronchiectasis, even in the absence of clinical features.

Feeding status of free-range scavenging chickens in different agro-climatic regions of India.

1. The aim of this study was to determine the nutritional status of backyard chickens reared under 4 different agro-climatic conditions (tropical, humid subtropical, cool-temperate and semi-arid) of India during different seasons of the year and to compare their nutritional status with those reared under confined feeding system (CFS). In each season, 15 adult and 15 grower chickens were slaughtered for collecting content of crop and gizzard to assess the nutritional status from different agro-climatic conditions.2. The dry matter content of crop and gizzard was higher (P < 0.01) in chickens reared under backyard conditions compared to those reared under CFS.3. Higher (P < 0.01) crop and gizzard contents were recorded during winter compared to the rainy season. The proportion of grains in the crop during summer and winter season was higher compared to rainy season. Further, there were more insects present in the crop during the rainy season and winter compared to summer.4. The proportion of insects was higher in chickens reared in humid, subtropical and cool temperate regions compared to semi-arid conditions. The consumption of grasses was higher in cool-temperate regions as compared to those reared under other areas.5. Crude protein (CP), Ca, P and gross energy (GE) content of the crop were higher in chickens reared under CFS compared to those reared in backyard systems and CP in the crop contents was lower in backyard chickens reared in humid subtropical and semi-arid regions compared to other areas. Similarly, lower (P < 0.01) Ca levels were seen in crop and gizzard contents from birds reared in semi-arid Rajasthan, and P and GE were lower in tropical wet and dry Telangana (TWT) compared to the chickens reared in the backyard conditions in other regions.6. Based on the results, the nutrient intake for birds utilising the scavenging feed resource base (SFRB) in the TWT region was below the requirements of backyard chickens from the other regions selected in the present study. GE and CP were critically deficient in the SFRB of the TWT region. Therefore, evolving location-specific supplementary diets using locally available feed resources to meet nutrient requirement of backyard chickens is required for optimising production potential.

Genotyping of viable Toxoplasma gondii from the first national survey of feral swine revealed evidence for sylvatic transmission cycle, and presence of highly virulent parasite genotypes.

Feral swine are known reservoirs of various pathogens, including Toxoplasma gondii. Here, we report the first national survey of viable T. gondii in feral swine in the USA. We paired serological surveys with parasite isolation and bioassay to evaluate the prevalence and genetic diversity of these parasites. From 2012-2017, sera and tissues from 1517 feral swine across the USA were collected for the isolation of viable T. gondii. Serum samples were initially screened for antibodies to T. gondii, and then the tissues of seropositive feral swine were bioassayed in mice. Antibodies were detected in 27.7% of feral swine tested by the modified agglutination test (1:25 or higher). Antibody positive rates increased significantly with age, with 10.1% of juveniles, 16.0% of sub-adults and 38.4% of adults testing seropositive. Myocardium (50 g) from 232 seropositive feral swine was digested in pepsin and bioassayed in mice. Viable T. gondii was isolated from 78 feral swine from 21 states. Twelve of the 78 isolates were pathogenic to outbred Swiss Webster mice and 76 of the 78 isolates could be propagated further in cell culture and were genotyped. For genotyping, deoxyribonucleic acid extracted from cell culture-derived tachyzoites was characterized by polymerase chain reaction restriction fragment length polymorphism using the genetic markers SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1 and Apico. Genotyping revealed 15 ToxoDB genotypes, including 43 isolates for genotype #5 (haplogroup 12), 11 isolates for #24, four isolates for #2 (haplogroup 3), two isolates for each of genotypes #3 (haplogroup 2), #4 (haplogroup 12), #216, #221, #289 and #297 and one isolate for each of genotypes #1 (haplogroup 2), #39, #66, #260, #261 and #299. Genotype #5 was the most frequently isolated, accounted for 57% (43/76) of the isolates, followed by #24, accounted for 14% (11/76). Genotypes #260, #289, #297 and #299 are new types. Genotype #289 was highly virulent to mice and originated from feral swine collected in Louisiana on the same day at the same location. Genotype #216 was previously demonstrated to be highly virulent to mice. Our results indicate moderate genetic diversity of T. gondii in feral swine in the USA, with the genotype #5 (haplogroup 12) dominant in the continental USA, whereas genotype #24 (10/14) was dominant in Hawaii, suggesting different population structures of the parasites among the two distinct geographical locations.

Cancer Pain, Anxiety, and Depression in Admitted Patients at a Tertiary Care Hospital: A Prospective Observational Study.

CONTEXT: Pain is the most common symptom in admitted cancer patients. The association between the severity of cancer pain and distress symptoms such as depression and anxiety is a subject of research. AIMS: The aim is to study the prevalence of pain, anxiety, and depression in admitted cancer patients and determine the association between pain and anxiety and depression at a tertiary cancer care institute. SETTINGS AND DESIGN: This was prospective observational study. SUBJECTS AND METHODS: We enrolled 393 cancer inpatients prospectively after written informed consent. Their disease details, presence, severity, and character of pain were recorded. Numerical Pain Scale was used for pain scores, self-reporting Hospital Anxiety and Depression Scale for anxiety and depression. STATISTICAL ANALYSIS USED: Normal data were analyzed with parametric, nonnormal with nonparametric methods, and categorical with the Chi-square test. RESULTS: The prevalence of moderate-to-severe pain was 41.5%, anxiety 20.3%, and depression 24.8%. Proportion of patients with anxiety and depression was 9.2% and 17.7% in patients with no pain; about 32.8% and 36.7% with severe pain, respectively (P < 0.000). In patients with no depression 6% had anxiety; with depression 44.9% had anxiety (P < 0.000). Odd's ratio to have anxiety and depression was 4.44 (95% confidence interval [CI] 2.0318-9.7024) and 2.92 (95% CI 1.5739-5.4186), respectively, in patients with pain as compared to no pain (P < 0.00). There was a positive correlation between pain, anxiety, and depression scores. CONCLUSIONS: There is strong association between the presence and severity of pain and distress symptoms such as anxiety and depression in admitted cancer patients.

Clinical and angiographic profiles and six months outcomes of smokers with acute ST segment elevation myocardial infarction undergoing primary percutaneous coronary angioplasty.

BACKGROUND: Outcomes of primary percutaneous coronary intervention (PCI) for acute STEMI (ST-segment elevation myocardial infarction) in smokers are expected to be better than non-smokers as for patients of acute STEMI with or without fibrinolytic therapy. OBJECTIVES: This comparative study was designed to evaluate the outcomes of primary PCI in patients with acute STEMI in smokers and non-smokers. Clinical and angiographic profile of the two groups was also compared. METHODS: Over duration of two year, a total of 150 consecutive patients of acute STEMI eligible for primary PCI were enrolled and constituted the two groups [Smokers (n=90), Non-smokers (n=60)] of the study population. There was no difference in procedure in two groups. RESULTS: In the present study of acute STEMI, current smokers were about a decade younger than non-smokers (p value=0.0002), majority were male (98.9% vs 56.6%) were male with a higher prevalence of hypertension and diabetes mellitus (61.67% vs 32.28% and 46.67% vs 14.44%, p=0.001) respectively. Smokers tended to have higher thrombus burden (p=0.06) but less multi vessel disease (p=0.028). Thirty day and six month mortality was non-significantly higher in smokers 4.66% vs 1.33% (p=0.261) and 5.33% vs 2.66% (p=NS) respectively. Rate of quitting smoking among smokers was 80.90% at 6 months. CONCLUSION: The study documents that smokers with acute STEMI have similar outcomes as compared to non smokers with higher thrombus burden and lesser non culprit artery involvement. Smokers present at much younger age emphasizing the role of smoking cessation for prevention of myocardial infarction.

Molecularly Confirmed Acute, Fatal Sarcocystis falcatula Infection in the Rainbow Lorikeets ( Trichoglossus moluccanus) at the Philadelphia Zoo.

The protozoan parasite Sarcocystis falcatula is an important cause of clinical disease in several avian intermediate hosts. The host range of S. falcatula is wide, and numerous outbreaks of acute sarcocystosis have been reported in passerine and psittacine birds in captivity in the Americas. Previous diagnosis was performed by serologic methods, light, and/or electron microscopic examinations with limited molecular confirmation. Here, we report histological and molecular diagnosis of acute, fatal S. falcatula infections in rainbow lorikeets ( Trichoglossus moluccanus) at the Philadelphia Zoo. Pulmonary sarcocystosis was suspected antemortem in 3 lorikeets (3-5 yr old); these birds died despite antiprotozoal therapy. The predominant lesion was pneumonia associated with S. falcatula-like schizonts in pulmonary vascular endothelium. The multilocus PCR-DNA sequencing ( 18S rDNA, 28S rDNA, ITS-1, and cox1) of frozen lung tissue confirmed S. falcatula infections in all 3 birds. Our results and previous studies suggest that acute pulmonary form of sarcocystosis is a major contributor of death to Old World psittacine birds.

Practical consensus recommendations on fertility preservation in patients with breast cancer.

Young women diagnosed with cancer today have a greater chance of long-term survival than ever before. Successful survivorship for this group of patients includes maintaining a high quality of life after a cancer diagnosis and treatment; however, lifesaving treatments such as chemotherapy, radiation, and surgery can impact survivors by impairing reproductive and endocrine health. Expert oncologists along with reproductive medicine specialists discuss fertility preservation options in this chapter since fertility preservation is becoming a priority for young women with breast cancer. This expert group used data from published literature, practical experience and opinion of a large group of academic oncologists to arrive at these practical consensus recommendations for the benefit of community oncologists.

Correlation of severity of chronic obstructive pulmonary disease with potential biomarkers.

BACKGROUND: Chronic obstructive pulmonary disease (COPD) is a non-specific inflammation, which involves the airways, lung parenchyma and pulmonary vessels. The inflammation causes the activation of inflammatory cells and the release of various inflammatory mediators such as interleukin-1 beta, interleukin-6 and tumor necrosis factor alpha (TNF-a). The present study was designed to assess the serum cytokines [Interleukin-1ß (IL-1ß), Interleukin-6 (IL-6), Tumor necrosis factor-α (TNF-α)] levels in chronic obstructive pulmonary disease (COPD) patients and they were correlated with severity of disease by spirometric measurements. MATERIALS AND METHODS: A total of 384 COPD patients and 50 healthy controls were enrolled in this study. The COPD patients were divided according to gold stages ie: mild, moderate, severe and very severe. 5 ml of venous blood samples were taken from all participants and it was collected in a test tube containing anticoagulant and then centrifuged at 3000 rpm for 10 min. Serum was separated and used to measure the amount of TNF-alpha, il-1beta, and IL-6. Spirometry was performed according to the criteria set by the Gold 2012 RESULTS: Tnf-α (pg/ml), IL-6 (pg/ml), IL-1ß (pg/ml) serum levels in COPD patients and healthy controls subjects were measured. Tnf-α and IL-6 serum levels were significantly (<0.001) higher in COPD patients compared to healthy control subjects. Likewise, IL-1 beta levels were also significantly (p-value = 0.022) higher in COPD patients compared to healthy control subjects. The distribution of Tnf-α, IL-6, IL-1ß (pg/ml) serum levels in COPD patients in relation to GOLD grading. There was a significant (p < 0.001) difference in the level of TNF-α, IL-6 and IL-1ß (pg/ml) among the severity of COPD. The posthoc analysis revealed that the TNF-α was significantly (p < 0.05) higher among the than mild, moderate, severe and very severe COPD patients. A similar observation was also found for IL-6. However, IL-6 was significantly (p < 0.05) higher among mild, moderate, severe and very severe COPD patients. There was significant (p = < 0.0001) difference in the level of IL-1ß in the different severity of COPD. The posthoc comparison test showed that IL-1ß levels were significantly (p < 0.05) higher among the mild, moderate, severe and very severe COPD patients. CONCLUSION: The present study signifies that the levels of TNF-α, IL-1ß, and IL-6 are directly proportional to the post-bronchodilator FEV1 percentage. Results provide population-based evidence that COPD is independently associated with low-grade systemic inflammation, with a different inflammatory pattern than that observed in healthy subjects. Overall, these results identify a novel systemic inflammatory COPD phenotype that may be the target of specific research and treatment.

Indigenous endophytic seed bacteria promote seedling development and defend against fungal disease in browntop millet (Urochloa ramosa L.).

AIMS: This study was conducted to investigate indigenous seed endophyte effects on browntop millet seedling development. We report that seed-inhabiting bacterial endophytes are responsible for promoting seedling development, including stimulation of root hair formation, increasing root and shoot length growth and increasing photosynthetic pigment content of seedlings. Bacterial endophytes also improved resistance of seedlings to disease. METHODS AND RESULTS: A total of four endophytic bacteria were isolated from surface-sterilized seeds and identified by 16S rDNA sequencing as Curtobacterium sp. (M1), Microbacterium sp. (M2), Methylobacterium sp. (M3) and Bacillus amyloliquefaciens (M4). Removal of bacteria with streptomycin treatment from the seeds compromised seedling growth and development. When endophytes were reinoculated onto seeds, seedlings recovered normal development. Strains M3 and M4 were found to be most potent in promoting growth of seedlings. Bacteria were found to produce auxin, solubilize phosphate and inhibit fungal pathogens. Significant protection of seedlings from Fusarium infection was found using strain M4 in microcosm assays. The antifungal lipopeptide genes for surfactin and iturin were detected in M4; culture extracts of M4 showed a positive drop collapse result for surfactins. CONCLUSIONS: This study demonstrates that browntop millet seeds vector indigenous endophytes that are responsible for modulation of seedling development and protection of seedlings from fungal disease. SIGNIFICANCE AND IMPACT OF THE STUDY: This study is significant and original in that it is the first report of seed-inhabiting endophytes of browntop millet that influence seedling development and function in defence against soilborne pathogens. This study suggests that conservation and management of seed-vectored endophytes may be important in development of more sustainable agricultural practices.

Sarcocystis strixi n. sp. from a Barred Owl ( Strix varia) Definitive Host and Interferon Gamma Gene Knockout Mice as Experimental Intermediate Host.

Here we report a new species of Sarcocystis with a barred owl ( Strix varia) as the natural definitive host and interferon gamma gene knockout (KO) mice as an experimental intermediate host. A barred owl submitted to the Carolina Raptor Center, Huntersville, North Carolina, was euthanized because of paralysis. Fully sporulated 12.5 × 9.9 µm sporocysts were found in intestinal scrapings from the owl. Sporocysts from the barred owl were orally fed to 4 laboratory-reared outbred Swiss Webster (SW) ( Mus musculus) and 8 KO mice. All mice remained asymptomatic. Microscopic sarcocysts were found in all 5 KO mice euthanized on day 32, 59, 120, 154, and 206 post-inoculation (PI), not in KO mice euthanized on day 4, 8, and 14 PI. Sarcocysts were not found in any SW mice euthanized on day 72, 120, 206, and 210 PI. Sarcocysts were microscopic, up to 70 µm wide. By light microscopy, the sarcocyst wall < 2 µm thick had undulating, flat to conical, protrusions of varying dimensions. Numerous sarcocysts were seen in the histological sections of tongue and skeletal muscles from the abdomen, limbs, and eye but not in the heart. By transmission electron microscopy, the sarcocyst wall was "type 1j." The ground substance layer (gs) was homogenous, up to 2 µm thick, with very fine granules, and a few vesicles concentrated toward the villar projections. No microtubules were seen in the gs. Longitudinally cut bradyzoites at 206 days PI were 7.8 × 2.2 µm. Based on molecular characterization using 18S rRNA, 28S rRNA, and cox1 genes and morphology of sarcocysts, the parasite in the present study was biologically and structurally different from species so far described, and we therefore propose a new species name, Sarcocystis strixi n. sp.

Isolation of viable Toxoplasma gondii, molecular characterization, and seroprevalence in elk (Cervus canadensis) in Pennsylvania, USA.

Toxoplasmosis is a worldwide zoonosis. The ingestion of uncooked/undercooked meat and consumption of water contaminated with Toxoplasma gondii oocysts excreted by felids are the main modes of transmission of this parasite. T. gondii has been reported in multiple cervid species; however, little is known of the parasite in North American elk (Cervus canadensis). In the present study, antibodies to T. gondii were detected in serum of wild elk from Pennsylvania collected during 2013-2016 by the modified agglutination test (MAT, cut-off 1:25); 221 of 317 (69.7%) had MAT titers of 1:25 in 19, 1:50 in 28, 1:100 in 34, and 1:200 or higher in 140. Thus most (44.1%) elk had relatively high titers. Seroprevalence was slightly higher in males (76.9%) than females (67.5%, not statistically significant, Chi-square tests, P<0.0001) and was higher in adults (76.5%) than yearlings (46.4%, Odds ratio 3.82; 95% CL 1.72-8.47; P=0.001) or calves (21.7%, Odds ratio 12.58; 95% CL 4.51-35.10; P<0.0001). Annual seroprevalence was relatively stable throughout the period tested and ranged from 66.6% to 72.2%. Of the 101 elk harvested in 2016, hearts were bioassayed from 20 elk and tongues were bioassayed from 56; all tongue samples were negative. Viable T. gondii was isolated from hearts of two female elk, one of these was a seronegative adult and the other was a calf with no serum available for testing. Both T. gondii isolates were cultivated in cell culture and DNA derived from tachyzoites was characterized using the PCR-RFLP markers including SAG1, SAG2 (5'- 3'SAG2 and altSAG2), SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico. One isolate belongs to ToxoDB PCR-RFLP genotype #2 and the other is genotype #5. Both genotypes are frequently identified in animals in North America.

Sarcocystis pantherophisi n. sp., from Eastern Rat Snakes (Pantherophis alleghaniensis) as Definitive Hosts and Interferon Gamma Gene Knockout Mice as Experimental Intermediate Hosts.

Here, we report a new species, Sarcocystis pantherophisi n. sp., with the Eastern rat snake (Pantherophis alleghaniensis) as natural definitive host and the interferon gamma gene knockout (KO) mouse as the experimental intermediate host. Sporocysts (n = 15) from intestinal contents of the snake were 10.8 × 8.9 µm. Sporocysts were orally infective to KO mice but not to laboratory-raised albino outbred house mice (Mus musculus). The interferon gamma KO mice developed schizont-associated neurological signs, and schizonts were cultivated in vitro from the brain. Mature sarcocysts were found in skeletal muscles of KO mice examined 41 days postinoculation (PI). Sarcocysts were slender, up to 70 µm wide and up to 3.5 mm long. By light microscopy, sarcocysts appeared thin-walled (<1 µm) without projections. By transmission electron microscopy, the sarcocyst wall was a variant of "type 1" (type 1i, new designation). The parasitophorous vacuolar membrane (pvm) had approximately 100-nm-wide × 100-nm-long bleb-like evaginations interspersed with 100-nm-wide × 650-nm-long elongated protrusions at irregular distances, and invaginations into the ground substance layer (gs) for a very short distance (6 nm). The gs was smooth, up to 500 nm thick, without tubules, and contained a few vesicles. Longitudinally cut bradyzoites at 54 days PI were banana-shaped, 7.8 × 2.2 µm (n = 5). Molecular characterization using 18S rRNA, 28S rRNA, ITS-1, and cox1 genes indicated a close relationship with other Sarcocystis parasites that have snake-rodent life cycles. The parasite in the present study was molecularly and biologically similar to a previously reported isolate (designated earlier as Sarcocystis sp. ex Pantherophis alleghaniensis) from P. alleghaniensis, and it was structurally different from other Sarcocystis species so far described.

Sarcocystis jamaicensis n. sp., from Red-Tailed Hawks (Buteo jamaicensis) Definitive Host and IFN-γ Gene Knockout Mice as Experimental Intermediate Host.

Here, we report a new species of Sarcocystis with red-tailed hawk (RTH, Buteo jamaicensis) as the natural definitive host and IFN-γ gene knockout (KO) mice as an experimental intermediate host in which sarcocysts form in muscle. Two RTHs submitted to the Carolina Raptor Center, Huntersville, North Carolina, were euthanized because they could not be rehabilitated and released. Fully sporulated 12.5 × 9.9-µm sized sporocysts were found in intestinal scrapings of both hawks. Sporocysts were orally fed to laboratory-reared outbred Swiss Webster mice (SW, Mus musculus) and also to KO mice. The sporocysts were infective for KO mice but not for SW mice. All SW mice remained asymptomatic, and neither schizonts nor sarcocysts were found in any SW mice euthanized on days 54, 77, 103 (n = 2) or 137 post-inoculation (PI). The KO mice developed neurological signs and were necropsied between 52 to 68 days PI. Schizonts/merozoites were found in all KO mice euthanized on days 52, 55 (n = 3), 59, 61 (n = 2), 66, and 68 PI and they were confined to the brain. The predominant lesion was meningoencephalitis characterized by perivascular cuffs, granulomas, and necrosis of the neural tissue. The schizonts/merozoites were located in neural tissue and were apparently extravascular. Brain homogenates from infected KO mice were infective to KO mice by subcutaneous inoculation and when seeded on to CV-1 cells. Microscopic sarcocysts were found in skeletal muscles of 5 of 8 KO mice euthanized between 55-61 days PI. Only a few sarcocysts were detected. Sarcocysts were microscopic, up to 3.5 mm long. When viewed with light microscopy, the sarcocyst wall appeared thin (<1 µm thick) and smooth. By transmission electron microscopy, the sarcocyst wall classified as "type 1j" (new designation). Molecular characterization using 18S rRNA, 28S rRNA, ITS-1, and cox1 genes revealed a close relationship with Sarcocystis microti and Sarcocystis glareoli; both species infect birds as definitive hosts. The parasite in the present study was biologically and molecularly different from species so far described in RTHs and we therefore propose a new species name, Sarcocystis jamaicensis n. sp.

Coexistence of hard palate carcinoma with tuberculosis: A rarity.

Coexistence of tuberculosis (TB) and palatal malignancy is a rare phenomenon and it has never been reported. Here, we present a case of hard palate carcinoma with TB in a 60-year-old male patient who was successfully managed by three pronged approaches by combining antitubercular therapy with chemotherapy and radiotherapy.