Aberrant autoantibody production is characteristic of systemic lupus erythematosus (SLE), but follicular regulatory T (TFR) cells potentially can suppress this abnormality. We investigate functional changes in TFR cells from SLE patients. Circulating TFR cells were collected from 19 SLE patients and 14 healthy controls (HC) to compare molecular expression and in vitro suppressive capacity of follicular helper T (TFH) cell proliferation. To reveal the stability of Foxp3 in TFR, pyrosequencing of conserved non-coding sequence (CNS) 2 at the Foxp3 gene locus was performed. We then tested IL-2 in SLE-TFR cells to check restoration of suppressor function. Programmed death-1 (PD-1) expression in SLE-TFR cells was positively correlated with anti-DNA antibody levels and disease activity. These cells had impaired suppressive function for TFH cells with decreased expression of suppression mediators forkhead box p3 (Foxp3), cytotoxic T-lymphocyte antigen 4 (CTLA4), and IL-2 receptor alpha (IL2Rα). Pyrosequencing identified hyper-methylation in CNS2 region of SLE-TFR cells comparing to HC. With In vitro IL-2 stimulation, PD-1 expression of TFR cells significantly decreased along with increased expression of Foxp3 and CTLA-4, especially in low-dose. Thus, SLE-TFR cells have functionally defective to TFH suppression, but low-dose IL-2 therapy might be useful to restore this ability.

The mechanisms of adaptive radiation with phenotypic diversification and further adaptive speciation have been becoming clearer through a number of studies. Natural selection is one of the primary factors that contribute to these mechanisms. It has been demonstrated that divergent natural selection acts on a certain trait in adaptive radiation. However, it is not often known how natural selection acts on the source of a diversified population, although it has been detected in phylogenetic studies. Our study demonstrates how selection acts on a trait in a source population of diversified population using the Japanese land snail Euhadra peliomphala simodae. This snail’s shell colour has diversified due to disruptive selection after migration from the mainland to islands. We used trail-camera traps to identify the cause of natural selection on both the mainland and an island. We then conducted a mark-recapture experiment on the mainland to detect natural selection and compare the shape and strength of it to previous study in an island. In total, we captured and marked around 1,700 snails, and some of them were preyed on by an unknown predator. The trail-camera traps showed that the predator is the large Japanese field mouse Apodemus speciosus, but this predation did not correlate with shell colour. A Bayesian approach showed that the stabilising selection from factors other than predation acted on shell colour. Our results suggest that natural selection was changed by migration, which could explain the ultimate cause of phenotypic diversification in adaptive radiation that was not due to predation.

Longevity is highly variable among animal species, and has coevolved with other of life-history traits, like body size and rates of reproduction. Telomeres, through their erosion over time, are one of the cell mechanisms that produce senescence at the cell level, and might even have an influence on the rate of ageing in whole organisms. However, uneroded telomeres are also risk factors of cell immortalization. The associations of telomere lengths, their rate of change, and life-history traits independent of body size are largely underexplored for birds. To test associations of life-history traits and telomere dynamics, we conducted a phylogenetic meta-analysis using studies of 53 species of birds. We restricted analyses to studies that applied the telomere restriction fragment length (TRF) method, and examined relationships between mean telomere length at the chick (Chick TL) and adult (Adult TL) stages, the mean rate of change in telomere length during life (TROC), and life-history traits. We examined 3 principal components of 12 life-history variables that represented: body size (PC1), the slow-fast continuum of pace-of-life (PC2) and post-fledging parental care (PC3). Phylogeny had at best a small-to-medium influence on Adult and Chick TL (r² = 0.190 and 0.138, respectively), but a substantial influence on TROC (r² = 0.688). Phylogeny strongly influenced life histories: PC1 (r² = 0.828), PC2 (0.838), and PC3 (0.613). Adult TL and Chick TL were poorly associated with the life-history variables. TROC, however, was negatively and moderate-to-strongly associated with PC2 (unadjusted r = -0.340; with phylogenetic correction, r = -0.490). Independent of body size, long-lived species with smaller clutches and slower embryonic rate of growth may exhibited less change in telomere length over their lifetimes. We suggest that telomere lengths may have diverged even among closely avian related species, yet telomere dynamics are strongly linked to the pace of life.

Background High power short duration (HPSD) radiofrequency-ablation (RFA) is highly efficient and safe while reducing procedure and RF time in pulmonary vein isolation (PVI). The QDot-catheter is a novel contact-force ablation catheter that allows automated flow and power adjustments depending on the local tissue temperature to maintain a target temperature during 90watts/4seconds lesions. We analysed intraprocedural data and periprocedural safety using the QDot-catheter in patients undergoing PVI for paroxysmal atrial fibrillation (PAF). Methods We included n=48 patients undergoing PVI with the QDot-catheter with a temperature controlled HPSD ablation mode with 90watts/4seconds (TC-HPSD). If focal reconnection occurred besides repeat ablation the ablation mode was changed to 50watts/15seconds (QMode). N=23 patients underwent cerebral MRI to detect silent cerebral lesions. Results Mean RF-time was 8.1+/-2.8min, procedure-duration was 84.5+/-30min. The overall maximal measured catheter-tip temperature was 52.0°C +/- 4.6°C, mean overall applied current was 871mA +/-44mA and over all applied energy was 316J +/-47J. The mean local impedance-drop was 12.1 +/- 2.4 Ohms. During Adenosine challenge n=14 (29%) patients showed dormant conduction. A total of n=24 steam pops were detected in n=18 patients (39.1%), while no pericardial tamponade occurred. No periprocedural thromboembolic complications occurred, while n=4 patients (17.4%) showed silent cerebral lesion. Conclusion TC-HPSD ablation with 90watts/4seconds using the QDot-catheter led to a reduction of procedure and RF time, while no major complications occurred. Despite optimized temperature control and power adjustment steam pops occurred in a rather high number of patients, while none of them lead to tamponade or to clinical or neurological deficits.

Context - DICER1-associated sarcomas are rare. These are currently described under a wide variety of appellations; morphologic characterizations in reported cases and sites of occurrence have also been disparate. Design – We aimed to review pediatric sarcomas associated with DICER1-mutation reported at our center, along with literature review, to identify histologic hallmarks for diagnosis. Results - A 12 year old girl with intracranial sarcoma mimicking mesenchymal chondrosarcoma, a 16 year old girl with broad ligament sarcoma mimicking fibrosarcoma and a 5 month old girl with vaginal sarcoma mimicking embryonal rhabdomyosarcoma showed DICER1-mutation. All three tumors though seemingly diverse, had an uncanny resemblance, comprising of a primitive mesenchyme-like spindle cell component with rhabdomyoblastic differentiation on immunohistochemistry. Primitive blastema, chondroid differentiation and foci of anaplasia mimicking pleuropulmonary blastoma histology were variably present. One case showed primitive neuroblastic differentiation. Though the constellation of features reported in literature is quite varied, rhabdomyoblastic differentiation has been ubiquitously reported in tumours across sites. Molecular testing showed gain of chromosome 8 in 2 cases. All 3 cases responded to alternating Vincristine, doxorubicin and cyclophosphamide / Cisplatin, etoposide and ifosfamide (VDC/PEI) backbone followed by maintenance chemotherapy. Conclusion - We highlight this morphologic hallmark of rhabdomyoblastic differentiation with or without chondroid differentiation, in primitive appearing pediatric sarcomas, especially of female genital tract and brain, which should raise a flag to test for DICER1 pathogenic variation. This is crucial in low / middle income countries where sequencing is not done routinely. Timely diagnosis can ensure appropriate treatment and implementation of surveillance protocols for those with germline mutation.

Aims: Limited information is available about the efficacy and adverse events of COVID-19 vaccines introduced into public use with Emergency Usage Licences without completing phase-3 trials. Data refer to healthy and mostly younger people, with lacking evidence about the protectivity. This study aimed determining seroconversion rates and levels specific to anti-S-RBD IgG and total anti-spike/anti-nucleocapsid IgM and IgG antibodies against SARS-CoV-2 after the administration of two doses of the inactivated SARS-CoV-2 vaccine in Turkey, comparing three types of antibodies. It was also aimed to assess short-term adverse events due to the vaccine. It is intended to answer the questions about efficacy and safety, which lack in phase trials, especially at community level. It is also aimed to collect data, which will form the basis for assessing whether antibodies are protective at different community settings. Methods: The study carried out in Turkey, comprised 282 healthcare workers who received two doses of the inactivated SARS-CoV-2 vaccine administered in two 3µg doses, 28 days apart. In day-28 after the second dose, anti-S-RBD IgG and total anti-spike/anti-nucleocapsid IgM and IgG antibodies against SARS-CoV-2 were detected by in-vitro chemiluminescence immunoassay. Results: At day 28 after the second dose, the seroconversion rates were found to be 92.9% for total anti-spike/anti-nucleocapsid IgG and 15.2% for IgM and 98.2% for anti-S-RBD IgG antibodies. The immunogenicity was affected by pre-vaccination natural COVID-19 history and age. The incidence of at least one adverse event was found as 29.8% after the first dose and 24.1% after the second dose, with the most common events being pain at the injection site, weakness, fatigue and headache. Conclusion: A high rate of seroconversion was observed with no serious adverse events. Prior natural COVID-19 history and age significantly contributed to strong immunogenicity. A booster dose appears required for older ages and individuals without immune response.

Cardiac lipoma is extremely rare. Here we presented a unique illustrative case of interatrial septal lipoma protruding into right atrial causing symptoms in a 54-year-old male. Echocardiogram and computed tomography showed a well-shaped, giant and fixed occupying located in interatrial septum and right atrium. The only manifestation was palpitation though the mass filled almost all atrium and compressed superior vena cava. The patient received resection of the large-sized lipoma sizing 87mm in diameter and weighing ~1000g. Pathological exam demonstrated mature lipocytes and substantiated the diagnosis of lipoma. The patient did well postoperatively and symptoms were resolved.

Background: The impact of physical activity (PA) on immune response is a hot topic in exercise immunology, but studies involving asthmatic children are scarce. We examine the level of PA and TV attendance (TVA) in asthmatic children to assess the role on asthma control and immune response to various stimulants. Methods: Weekly PA and daily TVA were obtained from questionnaires at inclusion of the PreDicta study. PBMC cultures were stimulated with phytohemagglutinin (PHA), R848, poly I:C and zymosan. Cytokines were measured and quantified in cell culture supernatants using luminometric multiplex immunofluorescence beads-based assay. Results: Asthmatic preschoolers showed significantly more TVA than their healthy peers (58.6% vs. 41.5% 1-3h daily and only 25.7% vs. 47.2% ≤ 1h daily). Poor asthma control was associated with less frequent PA (75% no or occasional activity in uncontrolled vs. 20% in controlled asthma; 25% ≥ 3x weekly vs. 62%). Asthmatics with increased PA exhibited elevated cytokine levels in response to stimulants, suggesting a readiness of circulating immune cells for type-1, -2 and -17 cytokine release compared to low-PA and high-TVA subjects. Low PA and high TVA were associated with increased proinflammatory cytokines. Proinflammatory cytokines were correlating with each other in in-vitro immune responses of asthmatic children, but not healthy controls. Conclusion: Asthmatic children show more sedentary behavior than healthy subjects, while poor asthma control leads to a decrease in PA. Asthmatic children profit from exercise, as elevated cytokine levels in stimulated conditions indicate an immune system prepared for a strong response in case of infection.

Epidermoid, dermoid and teratoid cysts are nonodontogenic benign lesions derived from the germinative epithelium, appearing any where of the body. In the oral cavity, these are uncommon and account for less than 0.01% of all the oral cysts. These cysts often remain asymptomatic for years however can become acutely symptomatic.

Cytokine Filter Application in COVID-19 Patients; Island of Hope for Crash and Burn Patients or Future Solution for All Septic Acute Respiratory Distress Syndrome (ARDS) PatientsAli Ghodsizad MD, PhD, FACC, FETCS, FACSThe COVID-19 pandemic crisis certainly has challenged the scientific community as well as entire world. While incidence numbers have decreased following expedited vaccination and precautions, still some patients present with COVID 19 related pneumonia and ARDS requiring Veno-Venous Extracorporeal Membrane Oxygenation (VV ECMO) support to survive.In COVID-19 patients a cytokine release syndrome concomitant with ARDS can lead to overwhelming clinical scenario. Geraci and colleagues report on their single center feasibility study looking at application of the CytosorbTM hemadsorption device which was used as a parallel circuit within the VV ECMO circuit.The authors give evidence for safety and feasibility of the CytosorbTM hemadsorption device use in 10 patients with COVID-19 related ARDS in combination with VV ECMO. They show a reduction of inflammatory markers and cytokines following hemadsorption treatment. The cytokine storm can cause a critical clinical picture of septic shock. Only under high vasopressor and inotropic support end organ perfusion can be maintained. The required invasive pressure ventilation with high PEEP and peak pressure can decrease the intrathoracic venous return further and contributes more to the shock physiology (1). We have to look at inspiring results from current single center experience carefully understanding the evolving nature of COVID-19 related ARDS. Other groups have used plasmapheresis and CVVH modifications in COVID-19 cases. Dominik et al have shown a significant benefit only using hemadsorption comparing to other used protocols (2).We have shown successful application of somatic stem cells in COVID-19 patients on VV ECMO at our center. We could observe a reduction of inflammatory markers following somatic stem cell application (3). COVID 19 ARDS patients who required VV ECMO support, underwent a Pulmonary Artery (PA)-catheter placement and allogenic human stem cell injection into the PA using the PA-catheter as part of our expanded access protocol (3,4).Brouwer and colleagues, another group working with hemadsorption, have actually shown reduced survival in patients undergoing hemadsorption therapy (5). Geraci and colleagues describe their overall VV ECMO survival for COVID 19 related respiratory failure to be > 90%. Others including our center have experienced a much lower survival in that patient population. So patient selection clearly is a key point. The results presented by Geraci and colleagues have to be taken as a pioneering step, which can help in ARDS and septic clinical scenarios with different pathology in future.

This clinical case represents an unusual manifestation of COVID-19 pneumonia which started as arterial hypertension as well as the development of post-viral inflammatory complications and long-covid syndrome. Several factors such as hypertension, bile duct disease, and age can affect the duration of COVID which can lead to long COVID

Biofilms are ubiquitous and notoriously difficult to eradicate and control, complicating human infections, industrial and agricultural biofouling. Current biofilm studies are commonly performed with the biofilm cultured on mono-interface and generally have neglected to consider more realistic biofilm, where diverse interfaces are involved. In our study, a reusable dual-chamber microreactor with interchangeable membranes was developed to establish multiple interfaces for biofilm culture and test. Protocol for culturing Pseudomonas aeruginosa (PAO1) on the air-liquid interface (ALI) and liquid-liquid interface (LLI) under static environmental conditions for 48h was optimized using this novel device. This study shows that LLI model biofilms are more susceptible to physical disruption compared to ALI model biofilm. SEM images revealed a unique ‘mushroom-shaped’ microcolonies morphological feature, which is more distinct on ALI biofilms than LLI. Furthermore, the study showed that ALI and LLI biofilms produced a similar amount of extracellular polymeric substances (EPS). As differences in biofilm structure and properties may lead to different outcomes when using the same eradication approaches, the antimicrobial effect of an antibiotic, Ciprofloxacin (CIP), was chosen to test the susceptibility of 48h-old ALI and LLI biofilms. Our results show that the minimum eradication concentration (MBCE) of CIP using our dual-chamber device reached 1600μg/ml, which is significantly higher than the conventional microtiter plate method (64μg/ml). The results highlight the importance of having a model that can closely mimic in-vivo conditions to develop more effective biofilm management strategies.

Objective: In this study, it was aimed to compare various parameters during surgery and postoperative cognitive functions in patients undergoing coronary artery bypass graft (CABG) surgery using different levels of perioperative oxygen. Methods: One hundred patients scheduled for elective CABG surgery were included in the study. The patients were divided into two groups according to arterial oxygen levels during surgery. In the normoxia group (NG) (n = 50), PaO2 was between ≥100mmHg and <180mmHg with at least 40% FiO2 mechanical ventilation (MV), and in the hyperoxia group (HG) (n = 50), PaO2 was ≥180 mmHg with 100% FiO2 MV. Hemodynamic parameters, peripheral oxygen saturation (SpO2), cerebral oxygen saturation (rSO2) measured from bilateral sensors, and blood gas values were recorded at the planned measurement times. Mini-mental state examination test (MMSE) was applied to the patients before and at the 12th, 24th hours; on the 1st, 3rd, 6th months after surgery. Results: Extubation time was found to be shorter in NG (p <0.05). Between the groups, rSO2 and mean arterial pressure (MAP) were found to be significantly lower in HG at the time of T4 measurement (p = 0.042, p = 0.038, respectively). MMSE values of the groups at the 1st, 3rd and 6th months were found to be significantly higher in NG (respectively p = 0.017, p = 0.014, p = 0.002). Conclusion: Hyperoxemia application during CABG may be associated with worse postoperative late-term cognitive functions. Keywords: cardiopulmonary bypass; hyperoxia; postoperative complications; cognitive function; spectroscopy, near-infrared.

In this report, We discuss a case of epidermoid cyst originating from the female vulva, which was completely excised by surgery. Pathologically confirmed as a vulva epidermoid cyst.

Salt stress causes changes in root apoplastic barriers, such as the endodermis and the exodermis, and these changes are associated with variation in abiotic stress tolerance. We explored variation in root apoplastic barrier traits, O2 consumption and root and shoot Na+ and K+ content in a diverse collection of commercial and wild barley accessions subjected to non-saline (control) and saline treatments. Lignin and suberin deposition in endo- and exo-dermal cell walls varied between the accessions and in response to salt treatments. Twenty-two wild barley accessions formed an exodermis in response to salt treatments, whereas the commercial barley cultivar Barke did not develop an obvious exodermis. Accessions with pronounced root barrier deposition tended to have lower O2 consumption relative to the accessions with less obvious barriers. Treatment with abscisic acid enhanced suberisation and lead to a pronounced formation of an exodermis in wild barley accessions, whereas treatment with an ethylene precursor had no obvious effect on suberisation. Principal component analysis revealed associations between suberin deposition, root and shoot Na+ and K+, and root respiration. The variation in root apoplastic barrier traits within the barley accessions represents a useful resource for future crop breeding to improve environmental stress tolerance.

Abstract Objectives: In patients with chronic rhinosinusitis (CRS), olfactory dysfunction (OD) is a common complaint. The focus of this research was to evaluate the effectiveness of olfactory training (OT) in patients with CRS after sinonasal surgery. Design : Prospective controlled trial. Participants: We enrolled 111 Korean CRS patients who underwent sinonasal surgery. Main Outcome Measures: At baseline and three months after starting OT, both participants were assessed by an olfactory function test and endoscopy. The Korean version of the Sniffin’ stick II (KVSS-II) was used to perform the olfactory function test. A visual analog scale and the Mini-Mental State Examination were used to assess nasal and psychological functions. Over the course of three months, five odorants were used in OT (rose, lemon, cinnamon, orange, and peach). Results: Over a 12-week duration, OT increased olfactory activity in nearly 62 percent of subjects than in non-OT subjects. The OT group had significantly higher olfactory outcomes for the total KVSS-II and identification scores than the non-OT group between the initial and follow-up assessments. The initial score influenced the degree of olfactory improvement after OT. Conclusions: OT patients exhibited significantly higher total KVSS-II scores compared with non-OT patients after sinonasal surgery; in particular, the odor identification score was different between the two groups. The results of this study show that a 12-week period of repeated short-term exposure to various odors could be useful in enhancing olfactory activity in patients who underwent sinonasal surgery for the improvement of sensory-neural olfactory impairment.

Elderly individuals exhibit unbalanced bone marrow (BM) effector T cell subset differentiation, such as increased T helper (Th)-1 and T cytotoxic (Tc)-1 cell frequencies, but the underlying mechanism still unclear. Endothelial cells (ECs) , which are instructive components of the BM microenvironment, exhibit the phenotype of semi-professional antigen-presenting cells and regulate T cell recruitment and activation. Thus, we compared the frequency and function of BM ECs, especially their capacity to regulate effector T cell subsets, between young and old healthy individuals, and explored the underlying mechanism of this immunomodulatory discrepancy. Although the young and old EC percentages were comparable, young ECs showed less reactive oxygen species and better migratory and tube-forming abilities than old ECs. Notably, young ECs regulated T cells to differentiate into fewer Th1 and Tc1 cells than old ECs. Reduced T cell activation molecules and inflammatory cytokines in young BM ECs may be the possible mechanism.

We study the focusing inhomogeneous nonlinear Schr\”odinger equation with inverse-square potential $$ i\partial_tu+\Delta u-\frac{a}{|x|^2}u+|x|^{-b}|u|^{2}u=0, $$ where $a>-\frac14$ and $0

Aim: Kidney transplant patients are amongst the high-risk groups for severe Covid 19. To date, no specific antiviral agent has been found uniformly effective against severe acute respiratory syndrome coronavirus 2 (SARS-CoV2). Favipiravir, the recommended drug by The Turkish Ministry of Health, was uniformly supplied to all patients diagnosed to have COVID 19 with a positive nasopharyngeal swap PCR test. The aim of our study was to retrospectively compare our kidney transplant recipients who developed COVID-19 infection started on Favipiravir to those who did not use Favipiravir for the clinical course of the disease with a special emphasis on the occurrence of side effects/adverse events. Methods: Thirty-seven consecutive KT recipients with a median age of 46 years and of whom 62.2% were women; 8 deceased /29 living related donor, with a 8.0 (5.5-12.5) years median duration of transplantation were included in the study. Results: Twenty-six (70.3%) patients received Favipiravir, 11(29.7%) did not. There was no statistical significance in baseline demographic, clinical and laboratory findings between the groups except that the Favipiravir group was older and had a higher requirement of oxygen treatment. There was no statistically significant difference in the course and outcome of COVID-19 infection, in the occurrence of side effects/adverse events related to Favipiravir between the two groups. Laboratory data at baseline, day7 and 30 were also comparable between the groups. Conclusion: Although the efficacy of Favipiravir in the treatment of COVID-19 infection is currently controversial, Favipiravir can safely be used in kidney transplant patients.

Supporting InformationYee-Fun Lim,* Chee Koon Ng, US Vaitesswar, Kedar Hippalgaonkar*

Author Information:

A 79-year-old man was referred for severe cardiac decompensation. Chest radiography showed severe pulmonary edema, and transesophageal echocardiography revealed a large quantity of vegetation on all aortic valve leaflets with severe aortic valve regurgitation, heterogeneous cavities adjacent to the aortic annulus, and ventricular septal rupture. We performed thorough and extensive debridement of the aortic root; including the infected ventricular septum, reconstructed the ventricular septum and aortic root using autologous and bovine pericardial patches; and placed a bioprosthetic stented valve. The patient was discharged without any complications, and without recurrence of the endocarditis in the four years post-surgery.

Background and purpose The incretin hormone, glucose-dependent insulinotropic polypeptide (GIP), secreted by the enteroendocrine K-cells in the proximal intestine, may regulate lipid metabolism and adiposity but its exact role in these processes is unclear. Experimental approach We characterized in vitro and in vivo antagonistic properties of a novel GIP analogue, mGIPAnt-1. We further assessed the in vivo pharmacokinetic profile of this antagonist, as well as its ability to affect high-fat diet (HFD)-induced body weight gain in ovariectomized mice during an 8-week treatment period. Key results mGIPAnt-1 showed competitive antagonistic properties to the GIP receptor (GIPR) in vitro as it inhibited GIP-induced cAMP accumulation in COS-7 cells. Furthermore, mGIPAnt-1 was capable of inhibiting GIP-induced glucoregulatory and insulinotropic effects in vivo and has a favourable pharmacokinetic profile with a half-life of 7.2 hours in C57Bl6 female mice. Finally, sub-chronic treatment with mGIPAnt-1 in ovariectomized HFD mice resulted in a reduction of body weight and fat mass. Conclusion and Implications mGIPAnt-1 successfully inhibited acute GIP-induced effects in vitro and in vivo and sub-chronically induces resistance to HFD-induced weight gain in ovariectomized mice. Our results support the development of GIP antagonists for the therapy of obesity.

Microbial diversity and community function are related, and can be highly specialized in different gut regions. The cloacal microbiome of Sceloporus virgatus provides antifungal protection to eggshells during oviposition – a specialized function that suggests a specialized microbial composition. Here, we describe the S. virgatus cloacal microbiome from tissue and swab samples, and compare it to tissue samples from the gastrointestinal (GI) tract and oviduct, adding to the growing body of evidence of microbiome localization in reptiles. We further assessed whether common methods of microbial sampling – cloacal swabs and feces – provide accurate representations of these microbial communities and whether feces might “seed” the cloacal microbiome or impact the accuracy of cloacal swab sampling. We found that different regions of the gut had unique microbial community structures. The cloacal community, in particular, showed extreme specialization averaging 99% Proteobacteria (Phylum) and 83% Enterobacteriacaea (Family). Cloacal swabs recovered communities similar to that of lower intestine and cloacal tissues, but fecal samples had much higher diversity and a distinct composition (62% Firmicutes and 39% Lachnospiraceae) relative to all gut regions. Finally, we found that feces and cloacal swabs recover different communities, but cloacal swabs may be contaminated with fecal matter if taken immediately after defecation. These results serve as a caution against the assumption that fecal samples provide an accurate representation of the gut, and that although cloacal swabs can reflect a portion of the lower GI tract microbiome, they may also result in a mixed community of gut and fecal microbes.