C1q nephropathy is a rare glomerular disease with characteristic mesangial C1q deposition noted on immunofluorescence microscopy. It is histologically defined and poorly understood. Light microscopic features are heterogeneous and comprise minimal change disease (MCD), focal segmental glomerulosclerosis (FSGS), and proliferative glomerulonephritis. Clinical presentation is also diverse, and ranges from asymptomatic hematuria or proteinuria to frank nephritic or nephrotic syndrome in both children and adults. Hypertension and renal insufficiency at the time of diagnosis are common findings. Optimal treatment is not clear and is usually guided by the underlying light microscopic lesion. Corticosteroids are the mainstay of treatment, with immunosuppressive agents reserved for steroid resistant cases. The presence of nephrotic syndrome and FSGS appear to predict adverse outcomes as opposed to favorable outcomes in those with MCD. Further research is needed to establish C1q nephropathy as a universally recognized distinct clinical entity. In this paper, we discuss the current understanding of pathogenesis, histopathology, clinical features, therapeutic options, and outcomes of C1q nephropathy. C1q nephropathy is a rare form of glomerulopathy first described as a distinct clinic-pathological entity by Jennette and Hipp in 1985 , 3. It uComplements are a heterogeneous group of 40 proteins circulating in the blood stream. They get activated by specific molecules like autoantibodies, immune complexes , carbohydrate molecules in the surface of microorganisms (the MB lectin pathway), or a low grade spontaneous activation called alternate pathway . They plThe pathogenesis of C1q nephropathy is still not clear. Specialized C1q receptors which help in the binding of immune complexes are found in the mesangial cells of the kidneys . The detThe histological patterns of C1q nephropathy are heterogeneous as outlined below.C1q nephropathy could be broadly classified into two subtypes on the basis of light microscopy: (1) MCD/FSGS group and (2) immune complex mediated proliferative glomerulonephritis (GN) group. The latter is an umbrella group for several morphological appearances including focal or diffuse mesangial proliferative GN, membranous GN, and membranoproliferative GN. In addition, the FSGS group has three variants, namely, collapsing, cellular, and “not otherwise specified” variants . The proImmunofluorescence microscopy is more specific than light microscopy. The mainstay of immunofluorescence microscopy is the use of antisera against immunoglobulins or compliment components or even proteins like albumin and fibrinogens. The pattern of staining such as granular, linear, mesangial, or capillary pattern as well as the anatomical location of the staining all would aid in making the specific diagnosis in a renal biopsy. Antiserum against C1q (prepared from goat) is more specific and stains the C1q fragment of the complement component C1. Staining for C1q is evident in all cases of C1q nephropathy, either in dominant or codominant fashion, mainly in the mesangium . ImmunogDiagnostic confirmation of C1q nephropathy is arrived at when amorphous electron dense deposits are demonstrated in the mesangium ± glomerular capillary wall. Podocyte injury can also be noted . These dC1q nephropathy is rare, with prevalence ranging from 0.2 to 2.5%  in biopsDue to its not-well-understood pathophysiology and varied clinical presentation, C1q nephropathy poses a management challenge. Early specialist consultation is essential. There are no randomized controlled trials that have evaluated the treatment of this condition. Current therapy involves treating the underlying light microscopic lesion, and outcomes vary accordingly. Immunosuppression, commonly in the form of corticosteroids, remains the mainstay of treatment. In steroid resistant cases, pulsed methylprednisolone, cyclophosphamide, azathioprine, Cyclosporine, mycophenolate, and tacrolimus have all been tried separately or in combination therapies with steroids with good response. Rituximab, a monoclonal antibody to CD20, has been used in a couple of patients who failed to respond to steroids with some promising results—one of them achieved normalization of renal function and the other avoided hemodialysis [As may be expected, patients with MCD have favorable outcomes compared with those with FSGS. In particular, those presenting with nephrotic syndrome and FSGS often show poor response to corticosteroid treatment . Even inOf 8909 native kidney biopsies processed between 1994 and 2002 at Columbia University in New York, 19 were retrospectively identified to have C1q nephropathy. Sixteen were available for follow-up (mean 27.1 months) of which 12 had received immunosuppressive therapy. Twelve (75%) of them had stable renal function, and 4 (25%) had progressive renal insufficiency. Seven out of 13 patients with proteinuria had partial or complete remission with or without immunosuppressive therapy. Two patients, both of whom had collapsing variant of FSGS, progressed to ESRD with a median renal survival of 81 months [In the largest cohort of patients reported yet, a review of 4048 native kidney biopsies from 1985 to 2005 at the University of Ljubljana, Slovenia, revealed C1q nephropathy in 72 biopsies. Of the 11 patients in FSGS group, every one of them presenting with nephrotic syndrome, one-third progressed to ESRD during a mean of 2.9 years of follow-up. In contrast, 77% of the 27 patients with MCD had complete remission of their nephrotic syndrome. Of the 20 patients with proliferative glomerulonephritis, 75% presented as having chronic kidney disease. The majority had stable renal disease after follow-up irrespective of immunosuppressive therapy .In a Japanese study which reviewed 16,860 renal biopsies between 1975 and 2004, 61 biopsies were diagnosed to show C1q nephropathy. Mean duration of follow-up was 7.2 years. Three out of 8 patients with FSGS developed ESRD 8 to 15 years after biopsy. In both MCD and FSGS groups, relapse of nephrotic syndrome was common [A 2014 study at Great Ormond Street Hospital for Children in London reviewed all biopsies of patients who presented with proteinuria or nephrotic syndrome between 1991 and 2011 and found 35 cases of C1q nephropathy. Thirty children received steroids, and 53% of them were sensitive to therapy. The study found that children with C1q nephropathy of MCD subtype had similar remission rates at four years compared to MCD disease controls, despite more frequent relapses. The long term renal outcome was not significantly different. Most of those with other histological appearances  were resistant to steroid therapy, required second-line drugs, and failed to achieve complete remission [In another study, 2221 children aged 3 to 15 years underwent percutaneous biopsy between 1975 and 2002 and 30 of them had biopsy proven C1q nephropathy. Of them, 18 children were asymptomatic and the remaining 12 had nephrotic syndrome (NS). The asymptomatic children had more degree of hematuria and the children with NS had more proteinuria. Both the groups had MCD in light microscopy as the underlying diagnosis in the majority of their patients (73%) and the remaining ones had immune mediated GN or FSGN. All these children with NS were treated with prednisolone with or without Cyclosporine. Only 4 of the asymptomatic children received the steroid therapy and the rest of them received dipyridamole. The degree of proteinuria improved in both groups but degree of hematuria improved more in the asymptomatic group .Said et al. retrospectively analyzed 24 patients with C1q nephropathy who had renal allograft with the mean age of 31 years. None of these patents had C1q nephropathy or SLE in their native kidneys. These patients developed C1q nephropathy in their transplanted kidneys on an average of 37 months. The light microscopy showed no lesions in about a third of the patients. Almost half of them had their usual antirejection treatment, 4 of them had pulsed steroids, one required plasmapheresis with ACE inhibitor, and one patient was treated with ACE inhibitor alone. Another patient had tacrolimus toxicity and had the dose lowered. The clinical outcome of the remaining patients was not available. The authors concluded that C1q deposition in the allografts was a mere morphological pattern and would have no clinical significance in most patients .The following are large, single center series of cases which looked at outcomes in C1q nephropathy.Although three decades have elapsed since C1q nephropathy was first proposed as a distinct clinical entity, it remains poorly understood and controversial. Some authors suggest that it is part of the spectrum of FSGS/MCD. Many reports have described different clinical presentation, histopathology, response to therapy, and outcomes, suggesting that it may be a combination of disease groups than a single entity. However, all published case series have been from single center reviews and appear to be skewed in their demographic profiles. Moreover, the clinical utility of diagnosing C1q nephropathy is yet to be fully established. While there is reasonable evidence to predict poorer outcomes with the presence of FSGS with C1q nephropathy, there have not been any studies which specifically compare clinical characteristics and outcomes of FSGS patients with and without C1q deposition. Routine addition of C1q staining in renal biopsies is not part of current guidelines due to costs and availability involved. Until multicenter, randomized controlled trials are undertaken specifically for C1q nephropathy and results are known, treatment strategy will remain focused on underlying light microscopic pathology, with standard first-line therapy of corticosteroids and immunosuppressive agents in resistant cases.Features of C1q Nephropathy. Diagnostic criteria are as follows:characteristic deposition of C1q in renal mesangium in a dominant or codominant fashion,absence of clinical or immunological features of SLE,type 1 MPGN to be excluded.MCD/FSGS,proliferative glomerulonephritis.Histological subtypes are as follows:asymptomatic hematuria or subnephrotic proteinuria,nephritic syndrome or nephrotic proteinuria,hypertension,renal insufficiency.Common presenting features are as follows:corticosteroids as first line,immunosuppressive therapy in steroid nonresponders.Treatment:nephrotic syndrome and FSGS—unfavorable outcome,MCD—better outcome.Prognosis is as follows:


The involvement of high mobility group box-1 (HMGB1) in various inflammatory and autoimmune diseases has been documented but clinical trials on the contribution of this pro-inflammatory alarmin in children with juvenile idiopathic arthritis (JIA) and systemic lupus erythematosus (SLE) are basically absent. To address the presence of HMGB1 and a soluble receptor for advanced glycation end products (sRAGE) in different subtypes of JIA and additionally in children with SLE, we enrolled a consecutive sample of children harvested peripheral blood as well as synovial fluids (SF) at diagnosis and correlated it with ordinary acute-phase reactants and clinical markers.Serum and synovial fluids levels of HMGB1 and sRAGE in total of 144 children  were determined by ELISA.The children with JIA and those with SLE were characterised by significantly higher serum levels of HMGB1 and significantly lower sRAGE levels compared to the healthy controls. A positive correlation between serum HMGB1 and ESR, CRP, α2 globulin was found while serum sRAGE levels were inversely correlated with the same inflammatory markers in children with JIA. Additionally, high level of serum HMGB1 was related to hepatosplenomegaly or serositis in systemic onset JIA.The inverse relationship of the HMGB1 and its soluble receptor RAGE in the blood and SF indicates that inflammation triggered by alarmins may play a role in pathogenesis of JIA as well as SLE. HMGB1 may serve as an inflammatory marker and a potential target of biological therapy in these patients. Further studies need to show whether the determination of HMGB1 levels in patients with JIA can be a useful guideline for detecting disease activity. Juvenile idiopathic arthritis (JIA) is a heterogeneous group of arthritis diseases with the onset before 16 years of age and persistent for more than 6 weeks. Systemic JIA (sJIA) is quite distinct from oligoarticular or polyarticular subtypes with the prominence of systemic and inflammatory features . In the Although the pathogenesis of sJIA and cSLE remains unclear, a novel studies seem to point in the direction of innate, rather than adaptive immune response , 5. As sAccording to previous studies, extracellular HMGB1 can amplify inflammation and enhance immune responses by interacting with the receptor for advanced glycation end-products (RAGE) –20. AlthAlthough data coming from clinical studies on children are still very few in number, the presence of antibodies to HMGB1 in JIA patients is documented , 27. FurThe aim of the present study was to determine the presence of HMGB1 and sRAGE in sera and synovial fluids in children with the different subtypes of JIA and juvenile SLE, in order to assess their possible association to clinical and serum markers for inflammation.The study was designed as a prospective trial in which a total of 144 children were enrolled. Among 97 patients with newly diagnosed JIA, 35 children had polyarticular type, 34 oligoarticular, and 27 systemic type of the disease. All JIA patients fulfilled the ILAR diagnostic criteria . Further^9/l), red blood cells , platelets , hemoglobin (g/l), haematocrit (%). For patients with SLE autoantibodies, rheumatoid factor (RF), and C3 and C4 complement components levels were also analysed.All the children were patients of Division of Pediatric Rheumatology and Immunology, University Hospital Centre Zagreb between 2007 and 2012. The study was approved by the Ethics Committee of the Zagreb University School of Medicine and parental informed consent was obtained for each child. For all patients, the following routine laboratory tests were performed at the Department of Clinical Laboratory Diagnostics, University of Zagreb School of Medicine: erythrocyte sedimentation rate , C-reactive protein levels , α2-globulins (%), γ-globulins (%), white blood cells count , always between 9 and 10 a.m. Aliquots of the peripheral blood were centrifuged at 3500 rpm for 5 minutes at room temperature, after which the supernatant is aspirated and stored in kriotube  at -70°C until used. The serum levels of alarmins were determined with commercially available enzyme-linked immunosorbent assay (ELISA) kits: RAGE , USA) and HMGB1  according to the manufacturer’s protocol. In addition, levels of HMGB1 and sRAGE were determined by the same kits in synovial fluid of 21 patients with oligoarticular and 7 patients with polyarticular when there was an indication for evacuation of the synovial fluid.p-value <0.05 was considered as statistically significant. ROC curves were constructed with the help of pROC package for R statistical programming language [loess) was added [All statistical analyses were performed with R statistical programming language . Differelanguage . On scatas added . Levels Basic laboratory findings of the children with JIA, SLE and healthy controls recruited in the study are shown in Table During the initial examination, all the patients with systemic-onset JIA presented with spiking fevers and skin rash as well as high ESR, CRP and α2-globulins. Twelve JIA patients presented with arthritis, at disease presentation. The rest of the patients with sJIA developed arthritis later on while being hospitalised. Ten patients presented with lymphadenopathy, seven with hepatosplenomegaly and three with serositis. All the patients with SLE were antinuclear antibody (ANA) positive, 87% had double stranded DNA antibodies (anti-dsDNA), 63% anti-Smith (Sm), 25% anti-SSA, 19% anti-SSB, 26% anti-ribonucleoprotein (RNP) antibodies, 69% anti-histones, but none of the patients were positive for rheumatoid factor (RF). Six SLE patients were presented with constitutional symptoms , five with malar rash, four with renal involvement, and two with polyarthritis (data not show).Serum levels of HMGB1 were significantly higher (P < 0.001) in children with three major types of JIA and in children with SLE, compared to healthy controls  had significantly higher HMGB1 serum levels compared with patients without  corresponding features (P < 0.001). Next, in a group with SLE children, we had four patients presenting with lupus nephritis who also had higher levels of HMGB1 . The difference was significant (P < 0.003).At the same time, compared to the healthy children, the children with all subgroups of JIA as well as children with SLE were characterized by lower serum levels of sRAGE, but the difference was significant only in patients with systemic-onset of JIA and in children with SLE Table .The median concentrations of synovial fluid HMGB1 in patients with oligoarticular and polyarticular JIA  were higher than in serum . In addition, the synovial fluid level of sRAGE were lower in both oligoarticular JIA and polyarticular JIA groups  than in serum  , CRP  and α2-globulin  , and ESR .Additionally, in children with SLE no association was detected between HMGB1 or sRAGE to the commonly used laboratory tests, but there was statistically significant and positive correlation between sRAGE levels and C4 complement component . Among the study participants with systemic onset JIA, levels of HMGB1 were significantly higher in patients with hepatosplenomegaly and serositis compared to those without these clinical characteristics . The CRP levels of systemic JIA patients presenting with hepatosplenomegaly or serositis were also significantly higher but HMGB1 was more sensitive than CRP in detecting hepatosplenomegaly or serositis in these patients, as shown by receiver operating characteristics (ROC) curve assessment. The area under the curve (AUC) for HMGB1 was 0.9941 and for CRP was 0.7294 . At the optimal cut-off value of 19595 pg/ml for HMGB1, the diagnostic sensitivity was 94% and specificity was 100%. At a cut-off concentration of 136 mg/l CRP revealed a sensitivity of 64% and a specificity of 100% . It is well known that HMGB1 is released at the site of joint inflammation and that the injection of HMGB1 into a normal joint causes development of arthritis –45. In tWe believe that our description of an inverse relationship of increased serum levels of HMGB1 and decreased serum levels of sRAGE in blood samples taken at diagnosis indicates that inflammation triggered by alarmins plays a role in pathogenesis of JIA and juvenile SLE. Furthermore, the results presented here suggest that alarmin may be a potential therapeutic target for immunotherapy of JIA and SLE, but also suggest that HMGB1 may be a valuable laboratory biomarker. Further studies in longitudinally collected blood samples over time from patients may reveal whether HMGB1 could be a valuable biomarker even more sensitive and specific than other available indicators of inflammation.


The results of clone analyses based on the 16S rRNA gene led us to speculate the presence of a chemo-synthetic microbial ecosystem, where chemolithoautotrophic thermophiles, primarily the bacterial order Aquificales, function as primary producers using H2 or sulfur compounds as their energy source and CO2 as their carbon source, and the organic compounds synthesized by them support the growth of chemoheterotrophic thermophiles, such as members of the order Thermales and the family Desulfurococcaceae. In addition, the dominance of members of the bacterial genus Herbaspirillum in the high temperature bottom layer led us to speculate the temporal formation of mesophilic zones where they can also function as primary producing or nitrogen-fixing bacteria.Physicochemical characteristics and archaeal and bacterial community structures in an iron-rich coastal hydrothermal field, where the temperature of the most active hot spot reaches above 100°C, were investigated to obtain fundamental information on microbes inhabiting a coastal hydrothermal field. The environmental settings of the coastal hydrothermal field were similar in some degree to those of deep-sea hydrothermal environments because of its emission of H For example, the presence of shallow vents located in a few meters of water on Vulcano Island, Italy have been known for much longer than deep-sea systems due to their proximity to the shore and easier accessibility and also some microbes have been isolated from the vents  and two novel strains of virus infecting the type species of the genus Aeropyrum were successfully isolated  to obtain a snap-shot of the microbial communities inhabiting the Yamagawa coastal hydrothermal field.2 (100 kPa). The vial was closed tightly using a butyl rubber stopper and sealed with a screw cap. No reducing agent was added. Samples were stored at 4°C in the dark until used.The study site was a coastal hydrothermal field on the Yamagawa Beach  , a coarsin situ along the transect using a probe thermometer. Dissolved oxygen (DO), dissolved hydrogen sulfide, and pH of pore water were determined in situ using a CHEMets Kit for Dissolved Oxygen , a twin pH Compact pH Meter , and a VISOCOLOR ECO Sulphide kit , respectively. We could not determine DO, dissolved hydrogen sulfide, or pH of the pore water of the surface layer due to an insufficient amount of pore water.The vertical temperature profile was determined Gas samples were collected from the bottom layer of each point  using a 50 mL syringe equipped with a 30 cm length needle. Gas contents were analyzed using gas chromatography  equipped with a thermal conductivity detector and a SHINCARBON ST packed column . Argon was used as the carrier gas.Iron contents of the sediments were determined using the method described by Lovley and Phillips . This meBacteria and for Archaea, respectively. Sequences shorter than 300 bp were omitted from further analyses.Genomic DNA for clone libraries was extracted from approximately 10 g of each sediment sample using an UltraClean Mega Soil DNA Isolation Kit  and the obtained DNA (8 mL) was concentrated to a volume of 500 μL with distilled water by following the manufacturer’s instructions. Bacterial and archaeal 16S rRNA genes were amplified by PCR using primers Bac27F  and Bac9Archaea or Bacteria) or sequences classified into the incorrect domain  were removed. Each OTU was assigned to a phylum  or to a family (for Archaea) using the RDP classifier (e.g. Thaumarchaeota) were not classified by the RDP classifier.The rRNA gene sequences with ≥95% similarity were assigned to the same clone type  in both bacterial and archaeal libraries using the FastGroupII program . Also, aassifier  set at ahttp//www.ncbi.nlm.nih.gov/). The best hit sequence for each representative sequence was collected and used along with the sequences obtained in this study for the following phylogenetic analyses. In order to determine the phylogenetic positions of the representative sequences, sequences were aligned using the ClustalW program  were assessed using phylogeny-based metrics, UniFrac . SequencThe sequences reported in this study were deposited in the DDBJ database under DDBJ/EMBL/GenBank accession numbers AB812106–AB812548.Escherichia coli DH5α for the bacterial 16S rRNA gene, and Aeropyrum pernix K1T (= JCM 9820T) for the archaeal 16S rRNA gene.The bacterial and archaeal 16S rRNA genes in each sample were quantified by a quantitative fluorescent PCR method using the Thermal Cycler Dice Real Time System Single , and SYBR Premix Ex Taq (TaKaRa Bio) and SYBR Premix Ex Taq II (TaKaRa Bio) were used for quantitative detection of the bacterial 16S rRNA gene and archaeal 16S rRNA gene, respectively, following the manufacturer’s protocol as previously described , while tThe maximum temperature of the sediment in the bottom layer (approximately 30 cm beneath sediment surface) of HS6 was 104°C with an average temperature of 101°C . The tem2  and a modest amount of H2  was also detected. These anaerobic gasses were also detected from the bottom layer of the sediment at approximately 70°C . These gases decreased with increasing distance from HS6. Oxygen was not detected in the gas emitted from the bottom of HS6, while hot fluid from the same point contained a modest amount of dissolved oxygen (1 ppm). Despite vigorous steam emission, the gas contents of the surface layer of HS6 were similar to those of atmospheric air. Dissolved sulfide also decreased with increasing distance from the bottom of HS6  from the bottom of HS6 to the surrounding zones, while atmospheric air diffuses from the surface layer.A high amount of sulfide was dissolved in the hot fluid from the bottom of HS6 . Gas emim of HS6 . This inSandy sediments consist of volcanic rocks such as basalt, pyroxene, olivine, and pumice. These minerals are known to bear substantial amounts of iron and aluminum. The amount of Fe(III) in the surface sediments showed a clear declining trend with distance from the hot spot (15.7–6.5 μmol per g sediment), while those of bottom sediments showed an increasing trend (3.0–6.5 μmol per g sediment) .i.e. steep gradient of temperature, reducing power (H2 and sulfide) and CO2 emission from the subsurface, presence of anaerobic zone with high temperature, and the presence of abundant Fe(III). On the other hand, the steep gradient of oxygen discriminates HS6 from deep-sea hydrothermal vents, which is derived from anaerobic gas emission from the bottom layer and direct exposure to atmospheric air.Thus, HS6 included physicochemical settings similar to those of a deep-sea hydrothermal vent, Archaea and Bacteria, respectively) were investigated. Rarefaction analyses showed that bacterial communities are much more diverse than archaeal communities  of this environment  at HSB  and two novel strains of virus infecting the members of this genus  at HSB . Sequenceropyrum . In addiis genus , 39 ensumal vent  and frommal vent , respectal vents . The famodictium , 58, GeoGeogemma , Hyperthrthermus , 71, andt Spring , which iThaumarchaeota, a recently described archaeal phylum ubiquitously detected from various environments  in HSB  (4.8–30.0%)  in HSB . The seqpirillum . The spepirillum , 52, 64.pirillum . Contrarpirillum , 60. In ocedures . However sources , 37. Con sources , dominan8–30.0%) . Unlike bacteria . Thus, tAquificae and Deinococcus-Thermus, were detected from HSB and DSB . Like other genera in the order Thermales, the only described species of the genus Rhabdothermus, R. arcticus, is a heterotrophic thermophile  in the HSB.Our data showed the emission of HAquificae, function as primary producers using H2 or sulfur compounds as their energy source and CO2 as their carbon source. The organic compounds synthesized by them support the growth of thermophilic heterotrophs, so-called “chemo-synthetic microbial ecosystems.” Of course, as many mesophilic microbes were detected from the samples, these microbes may possibly serve as part of the energy and carbon sources for this ecosystem.These findings inferred the presence of a microbial ecosystem in HS6 based on the chemical energy source provided by HSB. Namely, chemolithoautotrophic thermophiles, mainly Pyrobaculum and most members of the archaeal family Pyrodictiaceae grow by Fe(III) reduction and also by reduction of sulfur compounds such as S0 and thiosulfate (Geobacter and Shewanella, were detected in this study. In fact, the isolation of an iron reducer affiliated to the phylum Chloroflexi, which is the first report of this ability in the phylum, was achieved using sandy sediment from this environment (Some strains of the archaeal genus osulfate , 24, 65.ironment . Since t


The bacterial and Archaeal community composition was characterized by applying 16S rRNA gene pyrosequencing directly to soil DNA from samples. Geostatistics tools were used to reveal the heterogeneous distribution of bacterial composition at this scale. Soil physical parameters and land management explained a significant amount of variation, suggesting that environmental selection is the major process shaping bacterial composition. All taxa systematically displayed also a heterogeneous and particular distribution patterns. Different relative influences of soil characteristics, land use and space were observed, depending on the taxa, implying that selection and spatial processes might be differentially but not exclusively involved for each bacterial phylum. Soil pH was a major factor determining the distribution of most of the bacterial taxa and especially the most important factor explaining the spatial patterns of α-Proteobacteria and Planctomycetes. Soil texture, organic carbon content and quality were more specific to a few number of taxa . Land management also influenced the distribution of bacterial taxa across the landscape and revealed different type of response to cropping intensity  according to phyla. Altogether, this study provided valuable clues about the ecological behavior of soil bacterial and archaeal taxa at an agricultural landscape scale and could be useful for developing sustainable strategies of land management.Even though recent studies have clarified the influence and hierarchy of environmental filters on bacterial community structure, those constraining bacterial populations variations remain unclear. In consequence, our ability to understand to ecological attributes of soil bacteria and to predict microbial community response to environmental stress is therefore limited. Here, we characterized the bacterial community composition and the various bacterial taxonomic groups constituting the community across an agricultural landscape of 12 km Soil physicochemical properties and land management characteristics were determined for each sample. Bacterial diversity was characterized by massive inventory of the 16S rRNA gene sequences amplified from soil DNA. In addition to demonstrating the heterogeneous and spatially structured distribution of microbial abundance and diversity across the landscape, variance partitioning revealed that bacterial richness is mainly driven by soil texture and pH whereas land management is a strong determinant of microbial abundance and bacterial evenness. Altogether this previous study demonstrated the relevance of the landscape scale for deciphering microbial distribution patterns and processes, and for evaluating the effects of land management strategies on soil microbial resources.In a previous study , the distribution of microbial abundance and bacterial community diversity  was investigated across a landscape of 12 kmIn the present study we focused on the distribution of the composition of bacterial and Archaeal communities and of the various taxonomic groups constituting the community across the landscape. The following questions were addressed: are all taxa heterogeneously distributed at this spatial scale? Do they exhibit the same patterns? Which drivers or ecological attributes characterize each bacterial and archeal taxon at this scale? To answer these questions, a geostatistical approach was used to map and describe the spatial variability of community structure and taxa, and a variance partitioning approach was applied to identify and rank the ecological attributes for each taxon. Spatial descriptors were also integrated into the analysis to better interpret their relative contributions to taxa variation across a landscape and to examine other neutral processes shaping bacterial and archeal taxa distribution.2) and intensive agricultural croplands (9.22 km2) mainly under winter crops  in rotation with late-sown crops (spring barley). The whole area is flat or slightly sloping, under a continental climate with a mean annual air temperature of 10.4°C and a mean annual rainfall of 762 mm (period 1968–2011). Land management practices were clustered into six categories .The study was carried out on a monitored landscape of 13 km² located in Burgundy, France  characterized by deciduous oak-hornbeam forests  were randomly collected from a 4 m3, clay, silt and sand) were carried out by the Laboratoire d'analyse des sols d'Arras of INRA (http://www.lille.inra.fr/las) as described in Dequiedt et al.  and R888 (5′-CCGYCAATTCMTTTRAGT-3′) in a nested PCR strategy to add the 10-bp multiplex identifier (MID) barcode as initially described by Plassart et al. . Equal aThe raw data sets are publicly available in the EBI database system (in the Short Read Archive) under project accession no. PRJEB5219.The generated sequences were subjected to bioinformatic analysis using the GnS-PIPE developed by the GenoSol platform  and initially described by Terrat et al. . After aFactoMineR package ; Perennial plant cover (three frequently mown) grasslands, three blackcurrant (Ribes nigrum) and one Miscanthus (Miscanthus giganteus), n = 7); Catch Crop ; Minimum tillage ; Mechanical hoeing  and Conventional tillage .The variability of soil physicochemical properties across the studied area was assessed by subjecting the data to principal component analysis (PCA). Land management practices over the entire landscape were summarized by performing a factor analysis for mixed data to define land management clusters using the P < 0.001) vectors harboring a correlation ≥0.20 relative to the two NMDS axes were represented.Differences in community structure between samples were visualized by applying the weighted UniFrac metric and Nonmetric multi-dimensional scaling (NMDS). Soil physicochemical parameters and the relative abundance of the most dominant bacterial and archeal phyla and Proteobacteria classes were incorporated into the analysis by vector fitting against the bi-plot ordination of community structure. Significance of the vectors was assayed by 999 permutations. Only the most significant , microbial community structure  and the relative abundance of the most discriminant bacterial and archeal phyla and Proteobacteria classes. As these variables did not follow the strictly required Gaussian distribution, they were first transformed using the nonparametric rank-order  transformation prior to considering the spatial correlations . In addition to the six retained physicochemical properties and the clusters summarizing land management intensity, space was characterized by using a Principal Coordinates of a Neighbour Matrix approach (PCNM). The PCNM method was applied to the geographic coordinates and yielded 76 PCNM, representing the multiple spatial scales that the sampling scheme could perceive (Ramette and Tiedje P < 0.001), thereby limiting overfitting and problems due to co-linear variables (Ramette http://www.r-project.org/) using the vegan package  and agricultural plots  and ranged from 1.74 to 174 and 0.835 to 14.6 g · kg−1, respectively (Table S1). Soil properties were spatially structured in patches ranging from 600 to 900 m (Table S2), which reflected both the distribution of land management categories and the variations in pedological patterns  or clayey (median 34.3%) with alkaline pH . Organic carbon and total nitrogen contents were highly correlated , allowing taxonomic identification of the major bacterial and archaeal groups constituting the community in each soil sample. The NMDS ordination of Weighted UniFrac distance between samples revealed significant variation in community composition between soil samples across the landscape , and silt content , whereas the main environmental parameters involved in discrimination on the second axis were soil organic carbon , CaCO3  and also silt contents   and also revealed the significant marginal effect of space in shaping community variations .More precisely, the db-RDA analysis revealed that physicochemical data, land management and space explained 73% of the variation in community composition. This analysis confirmed that soil physicochemical characteristics and land management practices strongly contributed to community variation , and contrasted with the communities located at the extreme West and at the East  of the landscape , γ-Proteobacteria (11.3%), Actinobacteria (11.2%), δ-Proteobacteria (10.8%), Bacteroidetes (8.4%), Acidobacteria (6.0%), and Firmicutes (5.5%). The bacterial and archaeal taxa involved in the bacterial community discrimination on the NMDS analysis were identified by vector fitting against the ordination plot. The main taxa explaining the community composition discrimination across this landscape were: on the first NMDS dimension, α-Proteobacteria , Verrucomicrobia , Nitrospirae  as well as δ-Proteobacteria , Chloroflexi , Bacteroidetes , Planctomycetes  on the second NMDS dimension. Forest samples were distinguished by a higher relative abundance of α-Proteobacteria, Verrucomicrobia, δ-Proteobacteria and Planctomycetes, and a lower relative abundance of Actinobacteria, Choloroflexi, Gemmatimonadetes, Firmicutes, Bacteroidetes, and Nitrospirae , no interpolated mapping was performed for these two bacterial taxa.The relative abundance of each bacterial phylum constituting the community on the krigged maps was interpolated by geostatistical approach. These maps evidenced a heterogeneous distribution of all the studied phylum, supported by the cross validation statistics (Table S2), with an effective patch range between 149 and 1147 m Fig.. As smalα-Proteobacteria, Bacteroidetes and Nitrospirae exhibited similar and “spotty” distributions, corroborated by autocorrelation ranges around 500 m, and low v-parameter values confirming raw spatial processes at smaller distances (Table S2). The maps of Fibrobacteres, Armatimonadetes, Gemmatimonadetes, Crenarchaeota, and β-proteobacteria across the landscape were patchier  and variations in bacterial and archaeal taxa belonging to Bacteroidetes, Firmicutes, Chloroflexi, Gemmatimonadetes, Thaumarchaeota, Crenarchaeota, and Fibrobacteres Fig.. On the obia Fig.. An unusobia Fig..Although spatial patterns of microbial diversity have been well documented from micro-scale , which partly matched with the distribution of land management types across the landscape. Fibrobacteres, Armatimonadetes, Gemmatimonadetes, Crenarchaeota, and β-Proteobaceria were distributed in larger patches (around 600 m), which matched both the distribution of land management clusters and physicochemical characteristics. Contrastingly, the spatial patterns of taxa including Chlorobi, Actinobacteria, Planctomycetes, and δ-Proteobacteria were characterized by an intermediate patch size (about 700 m) which matched with soil characteristics and geomorphology, especially in the case of the “Sans fond” river location. Finally, taxa belonging to Thaumarchaeota, Verrucomicrobia, Elusimicrobia, and Firmicutes exhibited larger patches across the landscape (about 1000 m), which also matched with variations in soil characteristics and geomorphology. These contrasting patterns suggest that different drivers contrastingly shape bacterial taxa across the landscape. It also suggests that bacterial taxa might be differentially influenced by neutral processes .By characterizing the distribution of bacterial and archaeal taxonomic groups at the landscape scale, we were able to compile, for the first time, original maps for the 19 most abundant phyla constituting the community. These maps revealed a heterogeneous and spatially structured distribution for all taxa except the Acidobacteria and α-Proteobacteria, Planctomycetes, Crenarchaeota, and Verrucomicrobia could reflect the weak impact of dispersal-mediated processes. This is in agreement with the size of the patch, which matches with physicochemical variability across the landscape.Variance partitioning analysis of bacterial and archaeal taxa variation revealed that soil physicochemical characteristics and land management mainly contributed in explaining the spatial distribution of 16 of the 19 taxa. This suggests that the main process shaping the distribution of bacterial and archaeal taxa across the landscape is environmental selection determined by physicochemical properties and land-use. Repeated reports of the strong influence of local soil environmental heterogeneity had led to the conclusion that selection was the only process shaping soil microbial communities . The acidophilic attributes of some genera belonging to α-Proteobacteria and Verrucomicrobia and the basophilic attributes of some genera belonging to Planctomycetes and δ-Proteobacteria are coherent with the correlation between these taxa and pH reported in recent studies . These observations confirmed recent studies which highlighted a greater relative abundance of Fibrobacteres, Bacteroidetes, and a lower relative abundance of α-Proteobacteria in agricultural soils as compared to forest ecosystems , to a large extent, but also from disturbance intensity arising from human activities. Even though our study did not directly demonstrate that the influence of space was exclusively due to dispersal limitation of the populations constituting the community, our data would support this hypothesis. In addition, a spatial approach was used to complete and define new ecological attributes for most of the taxa identified. Further investigations should now be devoted to the spatial patterns of fungal communities to fully depict the mechanisms and drivers of soil microbial biodiversity, and a more thorough analysis of the link with soil functioning.


To estimate genetic variability in exon 3 of caprine myostatin gene in Marwari goats.MSTN) gene (Genebank accession no. DQ167575) was used to amplify the exon 3 region of MSTN gene in Marwari goat. The genetic variability in exon 3 of MSTN gene in Marwari goat was assessed on 8% polyacrylamide gel electrophoresis to detect single strand conformation polymorphism (SSCP) pattern.A total of 120 blood samples from unrelated Marwari goats were randomly collected from different villages of Bikaner (Rajasthan), India. Genomic DNA was extracted from whole blood using blood DNA isolation kit (Himedia Ltd.) as per manufacturer’s protocol. The quality of extracted genomic DNA was checked on 0.8% agarose gel. Specifically designed a primer set for caprine myostatin (MSTN gene in Marwari goat showed two types of conformation patterns on 8% polyacrylamide gel. One of the patterns showed only two bands and was considered as genotype AA, whereas another pattern having an extra band was designated as genotype AB. The frequencies of AA and AB genotype for exon 3 region of MSTN gene were calculated as 0.90 and 0.10, respectively.The exon 3 of MSTN gene in Marwari goat through SSCP analysis. This information could be utilized in future breeding plan to exploit the unique characteristics of Marwari goat of Rajasthan.Low level of polymorphism was observed at exon 3 region of  MSTN) or growth and differentiation factor 8 is a member of the transforming growth factor-β superfamily that plays an important role in the regulation of muscle growth and meat quality [MSTN gene encode a glycoprotein that is expressed widely in skeletal muscles [MSTN gene plays a critical role in myogenic differentiation and results in failure of myoblast to differentiate into myotubes through inhibition of MyoD activity and expression via Smad3 [MSTN gene that inactivated its expression or produced a non-functional protein [MSTN gene in muscular development [MSTN gene in goat is limited [Marwari goat is a major meat breed of Rajasthan (India) that is well-adapted to the arid environment and can tolerate higher salt loads and requires less water. The breed is known for its faster growth and breeding efficiency . The uni quality . The thr muscles . MSTN geia Smad3 . The altia Smad3 . Dramati protein . A numbe protein ,8, and selopment . However limited .Polymerase chain reaction (PCR) based single-strand conformation polymorphism (SSCP) technique has emerged as a simple, efficient, and powerful tool to detect small genetic variation in amplified product even at single base pair. Routine electrophoresis could not detect change in single nucleotide in a particular sequence of double-stranded DNA as the physical properties of the double strands are almost identical for both alleles. In SSCP technique, the double-stranded DNA after denaturation undergoes three-dimensional folding and may assume a unique conformational state based on its DNA sequence. The difference in shape between two single-stranded DNA strands with different sequences can cause them to migrate differently on polyacrylamide gel, even though the number of nucleotides is same, which is, in fact, an application of SSCP .MSTN gene in Marwari goat using the PCR-SSCP technique.Therefore, the present study was carried out to detect genetic variation in exon 3 of All essential procedures of sample collection were performed strictly as specified by Institutional Ethical Committee with minimal stress to animals.Random blood samples (n=120) were collected from unrelated Marwari goats from the field units of AICRP on Goat Improvement Project functioning at the Department of Animal Genetics and Breeding, College of Veterinary and Animal Science, Bikaner.The study was conducted at the Molecular Genetics Laboratory of Department of Animal Genetics and Breeding, College of Veterinary and Animal Science, Bikaner, Rajasthan, India.About 2-3 ml of venous blood was collected from jugular vein of each animal in EDTA-containing vacutainer tubes under sterile conditions. The vials were shaken gently after blood collection to facilitate thorough mixing and then kept in ice box containing ice packs. The samples were transported to the laboratory immediately and processed on the same day to extract genomic DNA.MSTN gene was amplified using one set of primer designed from caprine MSTN gene sequence (Genebank accession no. DQ167575). The PCR reactions were carried out in a final volume of 25 µL containing 80 ng of DNA template, 0.1 µM of each primer, 10.0 mM dNTPs, 2.5 U of Taq DNA polymerase , 5 µL ×5 reaction buffer, 1.5 mM MgCl2, and 11 µL ddH2O. An initial denaturation at 95°C for 5 min was carried out and followed by 40 cycles of denaturation at 94°C for 45 s, annealing for 45 s (Genomic DNA was extracted from whole blood by Blood Genomic DNA Purification Kit (HI Media Pvt. Ltd.). The exon 3 of for 45 s , and extfor 45 s .MSTN gene was detected by SSCP analysis. Aliquot of 5 µL PCR products were mixed with 5 µL of denaturing ×2 gel loading dye and heated for 8 min at 95°C and finally chilled on ice for 7 min. 8% polyacrylamide gel electrophoresis (PAGE) was carried out with denatured amplified product to identify the different genotypic patterns. The gels were stained with ethidium bromide, analyzed under ultraviolet light, and documented by UVP gel-documentation system.The polymorphism in exon 3 of MSTN gene in Marwari goat revealed two different conformation patterns on polyacrylamide gel , and bone morphogenetic protein genes are few candidate genes that play an important role in bone formation, birth weight, weaning weight, body condition, and muscle growth.A major gene model suggests that few candidate genes are responsible for relatively large proportion of genetic variation and could be utilized as marker . Such maMSTN allele with higher birth weights and yearling weights in Piedmontese crossbred cattle. A double-muscling phenomenon was observed in Belgian Blue and Piedmontese cattle following mutation in coding region of MSTN gene.Myostatin protein acts as a negative regulator of skeletal muscle growth and keeps the skeletal musculature within appropriate proportions . The proMSTN gene were shown to have significant difference among goat breeds. It was reported that TTTTA deletion phenomenon occurred in MSTN gene was unique for goats when compared with sheep, cattle, water buffalo, domestic yak, pigs, and humans [The polymorphisms of d humans ,24-26 foMSTN gene in Marwari goat keeping in view the scanty information available for caprine MSTN gene. The present study in Marwari goat revealed low genetic diversity in exon 3 of MSTN gene which is in agreement with the similar study conducted in Saanen and Boer × Guanzhong goats by An et al. [MSTN gene. However, a similar type of study in Sanjabi and Zel sheep by Soufy et al. [et al. [MSTN gene that suggests the fixation of this locus in sheep. The results of the present study indicate that low level of polymorphism still exists in this meat type of breed. The inconsistency in the results may be ascribed to species and breed differences, population and sampling size, environmental factors, mating strategies, geographical position effect, and frequency distribution of genetic variants. The present study could provide a base for further investigation of caprine MSTN gene in Marwari goats and other native breeds using large number of samples.The present study focused on identification of possible genetic variants in exon 3 of n et al. , who alsy et al.  and Dehn [et al.  revealedMSTN gene in Marwari goat revealed low level of polymorphism. This information could be utilized in future breeding plan to exploit the genetic potential of Marwari goat.The PCR-SSCP analysis of exon 3 of GCG and JPK designed the work plan. JPK, K, and ASD collected and processed the blood samples. JPK, MA, and VKA carried out PCR, electrophoresis, and PAGE. GCG, JPK, and P compiled, tabulated, transformed, and analyzed the data. JPK and GCG interpreted the results. JPK, VKA, and K prepared the manuscript. All authors read and approved the final manuscript.


Alouatta sp.) are obligatory trichromats. In all other platyrrines, only females can be trichromats, while males are always dichromats, as determined by multiple behavioral, electrophysiological, and genetic studies. In addition to obligatory trichromacy, Alouatta has an unusual fovea, with substantially higher peak cone density in the foveal pit than every other diurnal anthropoid monkey (both platyrrhines and catarrhines) and great ape yet examined, including humans. In addition to documenting the general organization of the retinal ganglion cell layer in Alouatta, the distribution of cones is compared to retinal ganglion cells, to explore possible relationships between their atypical trichromacy and foveal specialization. The number and distribution of retinal ganglion cells and displaced amacrine cells were determined in six flat-mounted retinas from five Alouatta caraya. Ganglion cell density peaked at 0.5 mm between the fovea and optic nerve head, reaching 40,700–45,200 cells/mm2. Displaced amacrine cell density distribution peaked between 0.5–1.75 mm from the fovea, reaching mean values between 2,050–3,100 cells/mm2. The mean number of ganglion cells was 1,133,000±79,000 cells and the mean number of displaced amacrine cells was 537,000±61,800 cells, in retinas of mean area 641±62 mm2. Ganglion cell and displaced amacrine cell density distribution in the Alouatta retina was consistent with that observed among several species of diurnal Anthropoidea, both platyrrhines and catarrhines. The principal alteration in the Alouatta retina appears not to be in the number of any retinal cell class, but rather a marked gradient in cone density within the fovea, which could potentially support high chromatic acuity in a restricted central region.Unlike all other New World (platyrrine) monkeys, both male and female howler monkeys ( Platyrrhines (New World monkeys) differ from catarrhines  in the variety of color vision phenotypes they exhibit. In catarrhine species, Old World monkeys and great apes, both males and females are trichromats, because in both males and females, three different genes (two in the X-chromosome and one in chromosome 7) code three different opsins. These opsins show the appropriate separation of light absorption peaks, and each opsin is expressed in a separate cone class Aotus sp., is an exception, a monochromat with a single cone class Alouatta are regular trichromats and their color vision seems to be very similar to that of catarrhines In most platyrrhine species, by contrast, only a proportion of females are trichromats . The remaining females and all males are dichromats because there are only two coding genes for opsins, one in the X-chromosome and another in the chromosome 7. In males, with only one X-chromosome and homozygous females, the result is dichromacy, while in heterozygous females gene polymorphism permits trichromacy Even though their color vision phenotypes are quite different, catarrhines and platyrrhines are quite similar in most aspects of their retinal anatomy. They have similar classes of horizontal cells Alouatta retinal anatomy. Franco and colleagues were able to estimate the cone density for both retinas of one Alouatta and found that they had an extremely high cone density in the foveal pit, about 429,000 cones/mm2 and 357,000 cones/mm2, respectively, accompanied by a comparably reduced cone diameter 2Alouatta caraya . The results show that the neurons of the ganglion cell layer are distributed following roughly the same pattern observed in other platyrrhines and also in catarrhines with minor specific differences. The overall ratio of cones to retinal ganglion cells for the entire fovea and foveolar region is as expected for other diurnal primates, so an acuity benefit from the region of high maximum density in Alouatta would have to be compensated by an acuity decrement in neighboring regions.There are only few studies of All animal experiments were carried out in accordance with the National Institute of Health Guide for the Care and Use of Laboratory Animals , and were approved by the Ethical Research Committee for Animal Experiments of the Institute of Biological Sciences, Federal University of Pará (#MED004/2008).Alouatta caraya were obtained from the breeding colony of the Centro Nacional de Primatas – CENP . The animals were bred and kept in the CENP facilities until the day the experiment started. There, they were kept in housing conditions permitting social interaction with their conspecifics and submitted to an appropriate feeding regimen and drinking water at libitum. Animal housing conditions, feeding regimen, and health were supervised by the CENP veterinarian staff. The animals were kept in environmental enriched cages as recommended by CENP primatologists.Five adults, male 2 of total area. Each room pair housed 1 male and 5–6 female Alouatta. Houses and rooms were cleaned daily. They were washed with jets of water containing 10% hypochlorite. Animals were moved from one of the paired rooms to the other during cleaning. Clean water was provided for primates in 500 ml animal drinking bottles, two bottles per room. Bottles were refilled 3 times per day. Primate diet consisted of the following items: a) food pellets provided in plates, ad libitum; b) fruits, vegetables, once a day; cooked eggs, twice a week; diluted milk, twice a week. In addition, Alouatta were provided, daily with leaves from Embauba tree, which are part of the natural diet of these Amazonian primates. All animals housed in the CENP received continuous veterinary care which followed the following protocol: inspection in the first hour every day to observe each animal and to inspect the feeding plates and drinking bottles; thereafter regular inspection until the evening. There are also facilities for primate anesthesia and surgery. The facilities allow primate full veterinary assistance, including X-ray, ultrasound, and complete blood, urine, and stool clinical laboratory examination and testing. Each room is enriched with trunks, branches, and ropes to facilitate primate exercising and escape during disputes and force display. Platforms are available for resting and access for feeding.Each animal room measured 2.50 m×4.00 m×2.25 m, 10 m−1) and then transferred to the Biological Sciences Institute of the Federal University of Para. The animal's electrocardiogram (ECG) was then recorded with thoracic surface electrodes, amplifier, and oscilloscope . Then, the animals were euthanized with an overdose of barbiturate, Thionembutal , 35 mg/kg or higher. Death was assessed by cessation of electrocardiogram ECG activity.On the day of experiment, the animals were sedated with an intramuscular injection of ketamine . The two retinas of the fifth animal were used in this work only (AC 05L and AC 05R).Animals were initially deeply anesthetized with the following intravenous anesthetics mixture: Levomepromazine  0.2–0.4 mg/kg; Midazolan , 0.2–0.4 mg/kg; Ketamine Chloridrate , 10–15 mg/kg. Just before perfusion they received Sodium Heparine 5,000 IU/ml (Roche), 1 ml i.v., to prevent blood coagulation followed by an intravenous lethal dose of Thionembutal , 35 mg/kg or higher. ECG was continuously monitored to ensure adequate depth of anesthesia and analgesia. Death was assessed by cessation of ECG activity. They were then transcardially perfused with 0.9% phosphate buffered saline solution , 500 ml, followed by 10% formaldehyde in PBS, 2000 ml. The eyes were removed, the retinas were dissected, mounted on a gelatinized slide with the ganglion cells uppermost, exposed to formaldehyde vapor 60°C for 2–3 h, then stained by the method of Nissl. Briefly, the retinas were rehydrated, stained with 0.5% cresyl violet  50–60°C under microscopic control, dehydrated in graded alcohols, cleared in xylene, and coverslipped.Measurements of retinal area were performed just after retinal dissection and in the end of all histological procedures. The results were compared in order to estimate retinal shrinkage. Retinal contours were drawn using a photographic enlarger at ×5 magnification and digitized using a HP Photosmart C5180 multifunctional equipment and the HP Scan and Camera Wizard software . Retinal area was then measured using the AutoCAD 2007 software .The positions of the fovea and optic disk center were also marked in the retinal drawings and their distance measured before and after histological procedure. The results were used to estimate if any shrinkage occurred in the central regions of the retina or were restricted to retinal borders and cuts made to obtain the flat mount. In addition, the vertical and horizontal lengths of the optic disk were measured on the microscope using a calibrated eyepiece.The retinal whole mounts were used to count ganglion cells and displaced amacrine cells along 12 radial directions, at 30° intervals, extending from the fovea towards retinal borders. The 0° and 180° directions hemisected the retina in the nasotemporal dimension passing through the fovea and optic disk while the 90° and 270° directions hemisected the retina in the dorsoventral dimension and passed through the fovea. These two lines were perpendicular to each other at the fovea and were called, in this work, horizontal and vertical meridians, respectively. Several works performed in primates in our laboratories and other laboratories used these two meridians as references, making possible to compare their results 2 in the first 1.5 mm from the fovea, then increased to 9,604 µm2 from 1.75 up to 5 mm, and then increased again to 38,416 µm2 from 6 mm up to the retinal periphery. Results were recorded as cells/mm2 on enlarged maps of the flat mounted retina, at ×10 magnification for the entire retina and ×40 magnification for the central 3 mm.Counts were performed under microscopic observation, using an ×100 oil immersion objective and ×10 eyepieces. The fovea was taken as the coordinate origin and counts were made at regular intervals from the fovea towards retinal borders. To account for the rapid cell density change in the first millimeters of distance from the fovea, counts were made at 0.25 mm intervals in the first 3 mm and then at 1 mm intervals up to the retinal borders. In addition, the counting area was 1,537 µmAlouatta retina. The isodensity contours were plotted linking points with density corresponding to the isodensity contour and points located between densities higher and lower than that corresponding to the isodensity contours. The total number of ganglion cells and displaced amacrine cells were estimated by measuring the area between two isodensity contours, multiplying the area times the mean density value of the two isodensity contours, and adding all the resulting figures.From the density values of ganglion cells and displaced amacrine cells measured at each retinal location, enlarged isodensity maps were drawn to illustrate the density topography for such cells of the Statistical analysis of the results was performed using software BioEstat 5.0 Six retinal whole mounts were successfully stained by the method of Nissl using cresyl violet as stain. Alouatta retina mounted with the ganglion cell layer facing the observer and stained by the method of Nissl. As in other platyrrhines and catarrhines, cell bodies of ganglion cells and displaced amacrine cells were radially displaced from the foveal pit and then packed in the several strata of the foveal slope. In the foveal slope, cell body density was very high, decreasing towards retinal periphery in all directions.Cebus monkey, 1.8 mm temporal to the fovea. Ganglion cells were labeled by biotinyl-lysine  Cercopithecus aethiops, Cebus apella, and Aotus trivirgatus labeled by this procedure using Biocytin or horseradish peroxidase (HRP) was used to consolidate the criteria to identify and count ganglion cells and displaced amacrine cells in platyrrhines Macaca fascicularis, generously provided by Professor Alan Cowey  in which one optic tract had been sectioned 13 months previously. This retina had bee prepared as a wholemount and stained with cresyl violet We also had available to us a retina from one Alouatta retinal flat mount stained with cresyl violet. The retinal ganglion cell layer is shown at three increasing eccentricities located temporal to the fovea. Neurons were readily distinguishable from glial cells and endothelial cells by their morphology. Ganglion cells comprised a heterogeneous population of neurons and were distinguishable from other neurons of the ganglion cell layer by the abundant amount of Nissl substance in their cytoplasm. The large- to medium-sized ganglion cells had a conspicuous amount of Nissl substance in the cytoplasm, an eccentric nucleus, and a clear nucleolus. These cells were readily labeled with Biocytin or HRP. The smaller ganglion cells had eccentrically placed nuclei, small rim of cytoplasm with small amount of Nissl substance, and were retrogradely labeled with Biocytin or HRP. All the neuronal classes that were positively identified as ganglion cells by Biocytin or HRP retrograde labeling were not found in the ganglion cell layer after optic tract section.Other neurons of the retinal ganglion cell layer comprised displaced amacrine cells and displaced horizontal cells, the later relatively rare in comparison with the former 2 (N = 4) and 641±62 mm2 (N = 6), respectively. The mean areal shrinkage in the four retinas that were measured before and after histology was 4.8±4.3% (range: 1 to 11%). As shrinkage occurred only along retinal borders and cuts performed to flat the retinas, no corrections were applied for cell density estimates obtained by cell counting.Five retinas were counted over their entire extent. An additional retina was partially analyzed once counts were performed in the midperipheral and peripheral regions, for eccentricities higher than 2–3 mm of distance from fovea. Alouatta retina, respectively. Ganglion cell counts were converted to ganglion cells per square millimeter and eccentricity values were taken as distances from the foveal pit. Ganglion cell density means and standard deviations are shown in expanded plots for the central 3 mm of eccentricity (A and C) (N = 5) as well as for the entire range of eccentricities (B and D) (N = 6). Ganglion cell density reached maximum values in the foveal slope: 44,963±3,089, 42,191±2,063, 43,825±788, and 40,673±2,669 cells/mm2 at 0.5 mm nasal, temporal, dorsal, and ventral to the fovea, respectively. Towards the retinal periphery, ganglion cell density steeply declined to the very low values: 274±131, 146±111, 166±63, and 97±78 cells/m2 in the nasal, temporal, dorsal, and ventral periphery, respectively. Ganglion cell density was higher in the nasal than in the temporal side at similar eccentricities. The nasal/temporal ration increased from 1.06 at 0.5 mm to 6.8 at 12 mm of eccentricity. There was no consistent difference in the dorsal versus ventral ganglion cell density.Alouatta retinas. In every figure, the upper map shows isodensity contours for the whole retina, while the lower map shows isodensity contours for the central region at a larger magnification. Isodensity contours were drawn from cell density values estimated at every retinal location that was measured and were exhibited as if observing from the internal retinal side. Ganglion cell density was expressed in cells/mm2. A cross and a gray oval indicated the locations of the fovea and optic disk, respectively. N and T indicate the nasal and temporal directions along the horizontal meridian. In the first retina (AC 01LM), the ganglion cell isodensity contours were slightly elongated in the nasal direction . The ganglion cell isodensity contours were elongated in the nasal direction reflecting the higher ganglion cell density in the retinal nasal quadrant, a feature present in different degrees for all the six retinas. The “average” retina also had a small elongation of the central isodensity contours along the vertical meridian, reflecting the presence of this feature in half of the retinas.Alouatta retina, represented as described for retinal ganglion cells. Displaced amacrine cell density reached maximum values in the foveal slope at 0.5–0.75 mm of eccentricity: 2,907±227, 2,719±411, 3,062±317, and 2,905±355 cells/mm2 nasal, temporal, dorsal, and ventral to the fovea, respectively. Toward the retinal periphery, displaced amacrine cell density initially declined at a fast rate and more slowly further in the periphery, reaching between 500–550 cells/mm2. There was no consistent density difference in the nasal versus temporal or dorsal versus ventral quadrants.2. At the retinal periphery this total remained between 600–850 cells/mm2. In the central region of the retina, the displaced amacrine cells represented between 6% and 6.7% of the total of neurons, while in the peripheral region they represented between 67% and 84% of ganglion cell layer neurons.The total of ganglion cell layer neurons reached maximum values in the foveal slope at 0.5–0.75 mm of eccentricity, between 43,500–47,900 cells/mmAlouatta retinas AC 01LM and AC 02LM, respectively. Alouatta retina, using the AC 02LM retina as template. Maps were drawing as described above for the ganglion cells. Displaced amacrine cell density was expressed in cells/mm2. Displaced amacrine cell density distribution followed a pattern approximately circular but with considerable variation in the central isodensity contours.The number of ganglion cells and displaced amacrine cells were estimated by integration of the isodensity maps. Alouatta compared to other platyrrhines and its atypical fovea compared to all other diurnal Anthropoidea, could make the Alouatta's visual system a naturally-occurring test of hypotheses about the role of the fovea in visual acuity and color vision. Though developmental data may be hard to acquire for this primate, the degree of disparity in density of cone packing within its fovea may be a natural test of several hypotheses of the mechanisms of foveal production Measurement of the number and types of retinal neurons and photoreceptors, their distribution, the exact conformation of the eye, and the neuron number and volume of central visual system structures has long been used as a source of information about how visual systems may be specialized for particular niches, contrasted with what features are conserved across visual systems HomoMacacaCercopithecusPapioCebusSaimiriCallithrixAotusThe topography of ganglion cell density and displaced amacrine cell density have been estimated in the retinas of several species of catarrhines and platyrrhines, either using retinal whole mounts alone or combining information gathered from retinal whole mounts and retinal sections. These studies have encompassed several genera of catarrhines and platyrrhines, including 2, with the highest value observed in the retina of the Callithrix jacchus jacchusAotus trivirgatus, the peak of ganglion cell density is about one third lower than in diurnal platyrrhines and catarrhines , the diurnal dichromat Cebus apella  Aotus trivirgatus  Callithrix jacchus jacchusCallithrix has a retina substantially smaller than Alouatta, Cebus, and Aotus, measuring about 200 mm2 versus 600–700 mm2 in the other three platyrrhines. Ganglion cell density in Alouatta and Cebus are largely similar. The ganglion cell density in Callithrix is higher than in Cebus and Alouatta in the foveal slope, becomes lower for intermediate eccentricities, and similar to the other two diurnal platyrrhines for more peripheral eccentricities.Cebus and Callithrix than in Alouatta. The nasotemporal asymmetry is due to a horizontal specialization, the visual streak, superposed to a radial specialization, the fovea and area centralis. Visual streaks are found in the retinas of many mammals and are particularly well developed in lateral eyed mammals such as rabbit Papio Anubis) Macaca mulattaCercopithecus aethiops sabeusCebus apellaSaimiri sciureusCallithrix jacchus jacchusAlouatta are large and slow-moving monkeys eating mainly top canopy leaves, and at lesser extent fruits, buds, flowers, and nuts, carefully not eating too many leaves of certain species without stopping, as some contain poison toxins Alouatta species do prefer the main canopy and emergent levels, but some species, such as Alouatta caraya, living in less favorable environments, come to the ground and cross open areas between forest patches Cebus, Callithrix, and Alouatta than in the nocturnal Aotus.The nasotemporal asymmetry is larger in Alouatta follows the common pattern of retinal ganglion cell distribution found in all diurnal platyrrhines and catarrhines: a distinct fovea whose size is approximately constant, about 250–500 µm in diameter in different species Alouatta shows is a very high cone density in the central fovea, to be discussed.In conclusion, Macaca mulattaMacaca fascicularisCallithrix jacchus jacchus2 for the ganglion cells connected to the most central cones, producing ratio of 1 cone to 2 or more ganglion cells thought to be critical in the evolution of trichromacy.The complexity of the primate fovea, with the displacement of cone cell bodies and all other neurons, makes it difficult to measure how the ganglion cell sampling density changes as a function of eccentricity in perifoveal and foveal regions. Taking into account the changing densities of cones, cone pedicles, and ganglion cells in combination with the offsets introduced by lateral displacement of cone pedicles, bipolar cells, and ganglion cells in the foveal region, these calculations have been performed for Alouatta results in higher total cone numbers in its fovea compared to Cebus , which would allow more time for the compaction of the central cones. Alternatively, perhaps some feature of the altered opsins themselves in this radiation permits greater variation in cone morphology.At this point, we can only speculate whether the high cone density in the central fovea of Alouatta  and Aotus  Aotus, the diurnal Alouatta has only 1.2 times more displaced amacrine cells.The totals of displaced amacrine cells estimated in five Aotus has developed a fully nocturnal retina in recent evolution, while Alouatta has all the features of a well-developed diurnal retina Click here for additional data file.S1 ChecklistThe ARRIVE Guidelines Checklist.(DOC)Click here for additional data file.S1 DatasetGanglion cell density, displaced amacrine cell density, total number of ganglion cells, cumulative number of cones and ganglion cells.(ZIP)Click here for additional data file.


Cervical intradural disc herniation (IDH) is an extremely rare condition, comprising only 0.27% of all disc herniations. Three percent of IDHs occur in the cervical, 5% in the thoracic, and over 92% in the lumbar spinal canal. There have been a total of 31 cervical IDHs reported in the literature. The pathogenesis and imaging characteristics of IDH are not fully understood. A preoperative diagnosis is key to facilitating prompt intradural exploration in patients with ambivalent findings, as well as in preventing reoperation. The purpose of reporting our case is to remind clinicians to consider the possibility of cervical IDH during spinal manipulation therapy in patient with chronic neck pain.The patient signed informed consent for publication of this case report and any accompanying image. The ethical approval of this study was waived by the ethics committee of Chonbuk National University Hospital, because this study was case report and the number of patients was <3.A 32-year-old man was transferred our emergency department with progressive quadriparesis. He had no history of trauma, but had received physical therapy with spinal manipulation for chronic neck pain over the course of a month. The day prior, he had noticed neck pain and tingling in the bilateral upper and lower extremities during the manipulation procedure. The following day, he presented with bilateral weakness of all 4 extremities, which rendered him unable to walk. Neurological examination demonstrated a positive Hoffmann sign and ankle clonus bilaterally, hypoesthesia below the C5 dermatome, 3/5 strength in the bilateral upper extremities, and 2/5 strength in the lower extremities. This motor weakness was progressive, and he further complained of voiding difficulty.Urgent magnetic resonance imaging (MRI) of the cervical spine revealed large, central disc herniations at C4–C5 and C5–C6 that caused severe spinal cord compression and surrounding edema. We performed C4–C5–C6 anterior cervical discectomy and fusion.The patient's limb weakness improved rapidly within 1 day postoperatively, and he was discharged 4 weeks later. At his 12-month follow-up, the patient had recovered nearly full muscle power.We presented an extremely rare case of cervical IDH causing progressive quadriparesis after excessive spinal manipulation therapy. The presence of a “halo” and “Y-sign” were useful MRI markers for cervical IDH in this case. He had no history of trauma, but had received physical therapy with spinal manipulation for chronic neck pain over the course of a month. The day prior, he had noticed neck pain and tingling in the bilateral upper and lower extremities during the manipulation procedure. The following day, he presented with bilateral weakness of all 4 extremities, which rendered him unable to walk. Neurological examination demonstrated a positive Hoffmann sign and ankle clonus bilaterally at that time, although these reflexes were not checked before. He complained hypoesthesia below the C5 dermatome, 3/5 strength in the bilateral upper extremities, and 2/5 strength in the lower extremities. This motor weakness was progressive, and he further complained of voiding difficulty.Urgent magnetic resonance imaging (MRI) of the cervical spine revealed large, central disc herniations at C4–C5 and C5–C6 with severe spinal cord compression and surrounding edema Figure . Further6 reported the first case of intradural cervical disc rupture, 31 cases of this rare condition have since occurred in the literature. In our review of these 32 cases, including our new case, we found a male predominance (19 of 32 cases) in IDH incidence, and the mean age at presentation was 45.2 years. The lower cervical spine, a site of increased spinal mobility, was the most frequently affected, particularly at C5–C6 and C6–C7.2 The most common clinical manifestations of cervical IDH were Brown–Sequard syndrome, transverse myelopathy, and rarely radiculopathy.4 In our new case, the patient presented with rapidly progressive quadriparesis after spinal manipulation therapy.Since Marega7 suggesting that traumatic injury may cause chronic adhesions between the ligaments and dura; indeed, these adhesions were found intraoperatively in most cervical IDH cases. In our patient, disc protrusion and repeated spinal manipulation caused chronic inflammation and mechanical irritation of the adjacent dura, creating adhesions between the PLL and the dura. We posit that the spinal manipulation procedures caused transdural disc herniation that compressed the spinal cord. Furthermore, we found evidence of adhesions between the dura and PLL in our patient intraoperatively.The mechanism of cervical IDH remains unclear. Trauma has been considered the most important inciting factor in previous studies,8 performed a cadaveric study that found the dura to be firmly attached to the PLL in some cases. They hypothesized that congenital factors may have a role in the pathogenesis of cervical disc herniation.Yildizhan et al10 They suggested that PLL hypertrophy and ossification can cause chronic mechanical irritation to the dura, particularly if it is already congenitally attached to the ligament.Some authors have reported cases of cervical IDH in patients with ossification of PLL.11 suggested that the presence of epidural gas on computed tomography could be an indicator. Choi et al5 designated an abrupt discontinuity in the PLL “hawk-beak sign” on MRI. Recently, Borm et al12 described a “Halo” of cerebrospinal fluid (CSF) isointensity surrounding a herniated mass on sagittal T2-weighted imaging that could indicate intradural extension into the cervical spine. Similarly, Sasaji et al13 described a “Y-sign” in the lumbar spine. Because intradural extra-arachnoid disc herniation occurs between the dura and arachnoid, the arachnoid was peeled from the dura by the disc herniation. One line of the dura and arachnoid was divided into 2 lines of the dura and the arachnoid. The branch of the ventral dural lines appeared as “Y,” and they called it the “Y sign.” In our case, a “halo” of CSF isointensity surrounding the herniated disc was demonstrated in axial MR images (Figure Because it is difficult to identify dural sac perforation on MRI, there have been few reports on the imaging features of IDH. In 1998, Hidalgo-Ovejero et al14 Since the anterior approach directly accesses the affected disc, it facilitates removal of extruded disc material and dura repair.Surgical intervention is the definitive treatment for cervical IDH, and all patients in the literature underwent surgery. According to previous reports, anterior cervical discectomy and fusion is superior to posterior decompression in terms of operative procedure and outcome.We presented an extremely rare case of cervical IDH causing progressive quadriparesis after excessive spinal manipulation therapy. The presence of a “halo” and “Y-sign” were useful MRI markers for cervical IDH in this case.


Layer chickens are exposed to high risks of production losses and mortality with impact on farm profitability. The harsh tropical climate and severe disease outbreaks, poor biosecurity, sub-minimal vaccination and treatment protocols, poor management practices, poor chick quality, feed-associated causes, and unintended accidents oftentimes aggravate mortality and negatively affect egg production. The objectives of this study were to estimate the probability of survival and evaluate risk factors for death under different intensive housing conditions in a tropical climate, and to assess the production performance in the housing systems.P < 0. 001) respectively. The total mean feed consumption/bird/day was similar for the open sided and sealed pens but the mean feed quantity per egg produce was significantly lower in the sealed pens ((P < 0.005). Seasons differently impacted on mortality with the hot-dry season producing significantly higher risk of mortality (61 times) and reduced egg production. Other parameters also differed except the egg production during the cold-dry season. Layers in sealed pens appear to have higher probability of survival and the Kaplan-Meir survival curves differed for each pen; ≥78 weeks old layer have higher probability of survival compared with the younger chickens and the 19–38 weeks age category are at highest risk of death (P < 0.001). The hazard-ratio for mortality of layers raised in sealed pens was 0.568 (56.8%).Daily mean mortality percentages and egg production figures were significantly lower and higher in the sealed pens and open houses  are at higher risk of death due to stress of coming into production, management changes and diseases, critical implementation of protocols that will reduce death at this precarious period becomes mandatory. Whether older chickens’ better protection from death is associated with many prophylactic and metaphylactic regimen of medications/vaccination will need further investigation. Commercial egg-type poultry production is relatively low in northern Nigeria compared with the southern zones in view of a number of factors including but not limited to the climatic conditions which aggravate mortality and negatively affect egg production percentages. Other factors including severe disease outbreaks, poor biosecurity, sub-minimal vaccination and treatment protocols, poor management, practices, poor chick quality, feed-associated causes, and unintended errors and accidents similarly influence production . Laying A negative association has been established between mortalities and net profits associated with lower egg productions ,4, and aDiseases that have negatively impacted on production and increase mortality in layer hens include amongst others the following: Infectious bronchitis  ; NewcastE. coli infections and these can contribute between 1 – 20% mortality and egg production losses of up to 50% based on severity [Others include Fowl typhoid, fowl cholera, fowl pox, infectious laryngo-tracheitis, Marek’s disease, Mycoplasma infections, infectious coryza, egg prolapse, aflatoxicoses, necrotic enteritis and severity -23.Meanwhile, certain other highly fatal diseases with transboundary and trade limitation potentials like the highly pathogenic avian influenza particularly the H5 and H7 subtypes will cause between 80-100% mortality, with egg production down to near zero and a follow-up policy of depopulation.The housing system has been confirmed to have major impact on mortality and production based on previous studies ,25. In SP < 0.001).The dataset comprised of 2,783 rows of observations, which represents the cumulative number of days from the initial mortality records (January 2010) to the end of study (December 2010) for all the 8 pens. Although the pen sizes remained the same, the stocking density and mean number of laying birds per pen differed. Specifically, the mean number of laying birds per pen were 8,414 (11.21%), 7,874 (10.49%), 7,452 (9.93%), 7,561 (10.07%), 6,447 (8.59%), 12,107 (16.13%), 14,149 (18.85%), and 11,072 (14.75%) for pen 1, 2, 3, 4, 5, 6, 7 and 8 respectively. Significant differences exist in total numbers of birds per pen initially housed and the final numbers taken out of each pen (Table P < 0. 001). Contrastingly, the egg production figures were significantly higher in the sealed pens compared to the open-sided ones , the values obtained here exclude those birds removed due to ill health, poor performance and aggressive behaviour all of which are removed by culling. The mean mortality percentages were significantly lower in the sealed pens (0.07%) compared with the open houses (0.18%) , pen number (1–8) and age of layers (in weeks) as covariates, layers in sealed pens appear to have higher probability of survival Figures  and 5 coP-values obtained during the multivariable analysis support the huge size of dataset used in this study (Table P < 0.001). Probability of mortality during the hot-dry season was 61 times compared with the cold-dry season which supports the descriptive analyses in Tables All of the five predictors screened at the univariable level were taken forward for consideration in the multivariable Cox proportional hazard regression model. The small confidence intervals and low dy Table . The fin2 per hen under the tropical environment [2 or 69.8 inch2) recommended for the ISA Brown breed. The sealed house with environmental controlled buildings had significant higher production figures with lesser mortalities due to the fact that the chickens were kept in more clement conditions since weather conditions were automatically managed.The stocking density varied widely in the analysis based on the observation of same house types but different flock populations resident in the buildings. There was an underutilization of full capacity of the different houses on the farm. Though we cannot immediately assess whether the stocking density directly impact on the production efficiency of the chickens and total egg laid but we confirmed that it did not influence the risk of mortality. Guo et al.  had confironment . From ouThe hot-dry season presented with extremely higher mortality figures compared to the other months and the risk of death during this period was also very high. This season might impact negatively on production performance due to associated heat stress, waning immunity and feed qualities and quantities consumed during this period. Although the feed consumptions during this same period was slightly higher than for the other periods, the manager did indicated that more feed tends to be wasted in these months and more water were taken that in the colder and warmer months. The FAO has demonstrated that at a temperature above 28°C, egg production will significantly wane both in quantity and quality . During It should be noted that an environmental temperature between 25-40°C will cause the bird to pant and may lead to heat stroke and eventual death . The cliSince the sealed pen type with environmental controls appear to reduce the risk of death in layer birds by about half, it will become important to promote this housing type in sub-Saharan Africa where climatic conditions get extreme at certain period of the year.It is noteworthy to state that younger birds 19–38 weeks) are at higher risk of death compared with older chickens  are at higher risk of death compared with older chickens, due to stress of coming into production, management changes and diseases, critical implementation of management protocols that will reduce death through other means at this precarious period becomes mandatory. Effort should be intensified to distinctly identify disease-associated and production/management-associated deaths in layer chickens as reasons for spiked mortality in layer chickens may not always be associated with disease. This is important in order to reduce the burden of drug administration in production animals.The farm is located in Bakura , a low poultry density area of the Savannah region of northwest Nigeria, away from human residential areas. The distance of the farm from the nearest highway is 5 km and approximately 13 km away from the neighbouring town. Each pen contained adequate number of cages to accommodate the population of chickens per house. The farm has 12 pens, 6 of which are open-sided pens and the remaining 6 are sealed pens. These pens are in three rows: The first row has 4 open pens, the second row has 5 with 2 open pens and 3 sealed and the last row has 3 sealed pens. The temperature and relative humidity control in the open pens are influenced directly by the supervening climatic conditions while the sealed pens are environmentally controlled automatically by thermostat-controlled monitors. All the pens were equipped with automatic 3-tiers battery cages with facilities including feeders, nipple drinkers, litter conveyor belts beneath each tier and a feed silo. The birds (ISA Brown) were placed in the house before the onset of lay at 13–14 weeks and remained therein till end of lay (90 weeks) in open pens and (100 weeks) in sealed pens. This variation in end of lay age is taken as the farm’s standard. The chickens were dewormed at week 8 and 14 and were vaccinated against the following pathogens: Mareks disease, Newcastle disease, Infectious bursal disease, Fowl pox, Fowl typhoid and Egg drop syndrome. The lighting program, the farm strictly adheres to the ISA Brown management protocols. The birds had 16 hours (maximum) of photoperiod as at the time the egg production reached 50% till the end of lay. Body weight, day and night length were considered in the determination of photoperiod length. In 2010, a prospective cohort study was conducted between 1st January and 31st December using longitudinal data from apparently healthy layer chickens from this regional commercial poultry farm. This farm largely supplies eggs and culled layers in northwest Nigeria. As at the time of data collection, only 5 open and 3 sealed pens were fully functional, giving a total of 8 pens used for the study. All records of daily, weekly and monthly activities were obtained through a data retrieval system on each chicken in the pens. Follow-ups documents and pathology reports/mortality records on each bird were obtained where necessary to exclude ambiguous data and iatrogenic causes of death within the period of study (365 days). All records of daily mortalities were also evaluated by either of the 2 resident poultry veterinarians to confirm their authenticities.A set of predictor variables believed to influence production parameters and mortality in poultry were obtained through the literature search ,24,30-33In the study, the unit of study is the pen and mortality rate is based on the number of dead chicken per house per day divided by the number of birds housed per pen. All statistical analyses were performed using Stata® v. 10 . Continuous variables were analysed using descriptive statistics: minimum, maximum, mean and median values. Daily mortality counts were modeled using survival analysis. The Kaplan-Meier estimator was used to estimate the probability of survival and median survival time and the log-rank test was used to compare survival curves.P < 0.20 in the univariable model for death or were biologically plausible in layer mortality were considered for inclusion in the multivariable model. Multivariable analysis, adjusted for confounding factors, was used to determine the hazard ratio (HR) for death. Variables were retained in the multivariable model if likelihood-ratio-test P < 0.05 [Univariable Cox’s proportional hazard modelling was performed to identify potential risk/hazard factors from the five variables initially considered. Those variables that demonstrated a statistically significant association(s) at P < 0.05 . The bacP < 0.05  is shownZi ⋅ ω.While the random effects was in P-values and 95% CI. For each covariate, HR relative to the chosen reference category was calculated in order to show their relationships.It was expected that observations with equal value of a variable in the data that identifies the group are assumed to have shared frailty. Across groups, the frailties are assumed to be gamma-distributed latent random effects that affect the hazard multiplicatively, or, equivalently, the logarithm of the frailty enters the linear predictor as a random offset. The results for the Cox proportional hazard model were reported as HR with its corresponding Although the observational data does not involve the handling and manipulation of animals, we ensured that all the “Five Freedom of Webster”  normal behaviour, Freedom from fear and distress) were taken care of in the farm. Similarly, the project was approved by the Research Committee of the Faculty of Veterinary Medicine, Usmanu Danfodiyo University, Sokoto. We also obtained and got clearance from the management of the farm to proceed and publish the outcome of the research.


Unfortunately the patient's response to therapy was limited and she subsequently died. We aim to review neuroblastoma in the context of increased intracranial pressure and the limited data of neuroblastoma occurring in the adult population, along with proposed treatment options.Neuroblastoma is the third most commonly occurring malignancy of the pediatric population, although it is extremely rare in the adult population. In adults, neuroblastoma is often metastatic and portends an extremely poor overall survival. Our case report documents metastatic neuroblastoma occurring in a healthy 29-year-old woman whose course was complicated by an unusual presentation of elevated intracranial pressures. The patient was treated with systemic chemotherapy, I Neuroblastoma is the most commonly occurring extracranial solid tumor diagnosed in the pediatric population , 2. NineNeuroblastoma is a disease of the sympaticoadrenal neural crest cells and may occur in any part of the sympathetic nervous system. Symptoms of the disease vary and are dependent on tumor location, presence of metastatic disease, and paraneoplastic syndromes. Typical sites of involvement include the adrenal medulla, neck, chest, and pelvis . Nearly μL, hematocrit of 18%, platelets of 32,000/μL immature myeloid cells, and nucleated red cells consistent with a leukoerythroblastic smear. Initial computed tomography (CT) scans of the head, chest, abdomen, and pelvis was notable for a 5 cm soft tissue mass in the right pelvis, anterior to the sacrum with a lytic lesion in the left iliac crest (μg (nl 0–135 μg).A previously healthy 29-year-old female originally presented with blurred vision, headaches, paresthesias of the fingers, and several days of fatigue. A complete blood count revealed a white blood cell count of 4,000/ac crest . Further2 on day 1 and etoposide 80 mg/m2/day on days 1–3 for the presumed diagnosis of metastatic neuroendocrine carcinoma. However, bone marrow evaluation revealed 80% marrow involvement of neuroblastoma with little evidence for differentiation. At this point, she was referred to our institution for further evaluation. The outside pelvic mass biopsy was reviewed. Taking into account imaging and bone marrow findings, the pelvic mass was best classified as a composite (nodular) ganglioneuroblastoma. A repeat bone marrow biopsy performed at our institution demonstrated extensive involvement by metastatic neuroblastoma  cytology was negative for involvement by neuroblastoma.At our initial visit, the patient continued to complain of visual changes and headache. Immediate ophthalmology evaluation revealed significant bilateral papilledema. A lumbar puncture was performed with opening pressure of >55 cm HBrain MRI was notable for diffuse dural enhancement and thickening and restricted diffusion within the calvarium concerning for metastatic disease to the dura and calvarium, with absence of normal flow in the transverse sinuses secondary to dural compression. Intravenous acetazolamide and methylprednisolone were administered for elevated intracranial pressure with initial improvement in her visual symptoms. Her serum vanillylmandelic acid and homovanillic acid prior to subsequent treatment were 243 ng/mL (nl < 20) and 93 ng/mL (nl < 30), respectively. She commenced antineoplastic therapy with topotecan 1.2 mg/m²/dose and cyclophosphamide 400 mg/m²/dose on days 1–5. The patient was discharged post-therapy with a stable ophthalmologic exam.μL), given her degree of bone marrow infiltration with neuroblastoma. Following cycle two, she developed worsening headaches and vision changes, with MRI findings of increased dural enhancement and compression of the sagittal sinuses without development of venous thrombosis. A lumbar drain, later converted to a ventriculoperitoneal (VP) shunt, provided minimal symptomatic relief.She then received her second cycle of cyclophosphamide and topotecan despite thrombocytopenia  itself could not be excluded. Though CNS involvement was suggested by dural enhancement seen on MRI, repeat cerebrospinal fluid (CSF) sampling over a period of months was negative by cytology. CNS involvement is very uncommon in neuroblastoma, with a rate of 0.6% in one large case series . When thOur patient's primary disease appeared to arise from a lesion in the pelvis. In a series of 25 adult/adolescent patients, 16% had disease occurring in the pelvis, while 68% arose in the retroperitoneum or adrenal glands .This case represents the presentation of a common disease in an uncommon patient population. The majority of adult oncologists will not experience caring for a patient with neuroblastoma; however, this diagnosis should at least be considered in certain cases. Neuroblastoma may occur in similar sites to neuroendocrine tumors and should be considered particularly in patients of adolescent age and those with neurologic paraneoplastic symptoms. Furthermore, due to the rarity of these diseases, diagnostic evaluation by an experienced pathologist is warranted to distinguish between neuroendocrine tumors and the rare case of adult neuroblastoma.